TW201040274A - Rice transgenic event 17053 and methods of use thereof - Google Patents

Abstract

The present invention provides a transgenic rice event 17053 and plants, plant cells, seeds, plant parts, and commodity products derived from event 17053. The present invention also provides polynucleotides specific for event 17053 and plants, plant cells, seeds, plant parts, and commodity products comprising polynucleotides specific for event 17053. The invention also provides methods related to event 17053.

Description

201040274 VI. Description of the Invention: [Technical Field to Which the Invention Is Ascribed] The present invention relates to a genetically modified rice line Π053. Plants containing this line showed tolerance to glyphosate herbicides. The invention also relates to nucleic acid molecules, plants, plant parts, plant seeds, plant cells, agricultural products and methods related to line Π053. The present invention provides nucleotide molecules which are unique to this strain and which are combined with the insertion of gene-transformed DNA into the rice plant genome. This application claims the benefit of U.S. Provisional Application Serial No. 61/164,899, filed on March 30, 2009, which is hereby incorporated by reference. The sequence listings accompanying the application are incorporated herein by reference. [Prior Art]

Rice is an important crop in many parts of the world. The biotechnology method has been applied to rice to produce rice with improved properties. One of these improved properties is herbicide tolerance. The herbicide-tolerant transgenic gene is expressed in plants for the purpose of producing plants having the desired herbicide tolerance characteristics. The expression of the transgenic gene in plants may be due to the close proximity of the chromatin structure (eg, heterochromatin transcriptional regulatory elements (eg, enhancers) close to the integration site: the effect of the chromosomal location of the gene. A large number of individual plant transformation lines are selected to identify lines that have the best performance of the transgenic genes and therefore have the specific desired characteristics. For example, the number of transgenic genes that have been observed in plants can vary widely. There may also be differences in spatial or temporal patterns 'eg relative transgenic groups in various plant tissues 147377.doc 201040274 due to the amount of performance, ΓΛ ΓΛ μ / / 旎 旎 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 The transcriptional regulatory element is expected to be the first tenth of the body. For this reason, Kewei needs to produce hundreds to thousands of different gene-transferred lines and screened for 2 copies in these lines. Lines of gene expression and pattern are used for commercial purposes. The line with the amount or pattern of gene transfer can then be used for sexual hybridization using plant breeding methods. Transgenic genes infiltrate into other genetic backgrounds. These hybrid progeny will have the transcriptional gene expression of the original transformant: this can be used to ensure a robust gene in many different varieties that are appropriately adjusted for growth conditions in a particular region. SUMMARY OF THE INVENTION The present invention provides a genetically-transferred rice plant comprising line i7053 which exhibits a commercially acceptable tolerance to the application of a glyphosate herbicide, the representative seed being deposited under accession number PTA_9843 American Species Preservation Center (ATCC). The present invention also provides seeds, progeny, plant parts, cells, and commercial products comprising lines 17 to 53 meters. The present invention also provides novel DNA related to the genome containing the line 17053. Molecules and methods of using the same. The invention also provides methods of using the genetically modified rice line 1 7053, and plants comprising the lines and methods of producing glyphosate-tolerant rice.

The present invention provides DNA molecules associated with rice line 17053. The DNA molecules may comprise a nucleotide sequence representing or derived from: a junction sequence between a transgenic insertion sequence of rice line 1 7053 and a flanking gene dna and/or flanked by the inserted DNA The genomic DNA region, and/or the integrated gene-transforming DNA region flanked by the insertion site, and/or the integrated gene-transforming region of the cassette, and/or any of the regions 147377. Doc 201040274 The contiguous sequence of people. The present invention also provides a DNA molecule for use as a primer and probe for identifying rice line i7053. Rice plants, plant cells, plant parts, commodities, progeny and seeds comprising such molecules are also disclosed. The present invention provides methods, compositions and kits for detecting the presence of DNA derived from rice line 17053. The present invention provides a method for detecting a strain of 1 〇53, which is obtained by contacting a sample containing DNA with a primer set which is used for nucleic acid amplification reaction with the gene DNA of rice line 17053. Amplification of DNA of rice line 17053; performing nucleic acid amplification reaction to thereby generate amplified DNA; and detecting amplified DNA to achieve. The present invention also provides a method for detecting a line 17053 by contacting a sample containing DNA with a probe of the gene DNa of the rice line 17〇53 for the heterogeneous reaction day ττ′ and the rice line. 17053 is characterized by hybridization of DNA molecules; performing heterozygous reactions; and detecting hybridization of probes with DNA molecules. Kits comprising the methods and compositions of the present invention for use in detecting the presence of DN A from rice lines 17 〇 5 3 are also provided. 〇 The present invention provides a rice plant, seed, plant cell, progeny plant, plant part, or product derived from the plant, plant cell or seed, comprising line 17053. The present invention also provides a DNA molecule comprising a DNA sequence having a nucleotide sequence selected from the group consisting of SEq ID no: i_6 and complementary sequences and fragments thereof, or a DNA molecule having at least 90% sequence identity to SEQ ID NO: Rice plants, seeds, plant cells, progeny plants, plant parts or commodities. The invention also provides a rice plant, seed, plant cell, subunit comprising a line 17〇53 and comprising, for example, a DNA molecule comprising a DNA molecule comprising the amplified DNA molecule of SEQ ID NO: 1 and/or SEQ ID NO: 2, by DNA amplification. 147377.doc 201040274 A plant, plant part or commodity derived from the plant or seed.八本^ provides a method of controlling weeds in the field by planting the rice of the line 3, line 17053 and subsequently applying a herbicide scale herbicide capable of controlling the weeds without harming the plants containing the line 17053. °. The present invention provides a method for producing a rice plant and/or seed which is resistant to the application of a glyphosate-tolerant herbicide by using a plant containing about 7053 or containing _ID shame WSEQ ID N〇: 2 The second rice plant has sexual crosses' thereby producing seeds, planting the seeds to produce progeny plants, treating the progeny plants with glyphosate and selecting progeny plants resistant to glyphosate. Narrow methods can also include selfing the selected progeny plants to produce a plurality of sub-secondary plants and selecting glyphosate-tolerant plants from the second generation plants. The methods may also include sexually crossing the selected progeny plant with another rice plant to produce seeds, planting the seed to produce the second generation plant, treating the second generation plant with glyphosate and selecting the second generation tolerant to glyphosate. plant. The present invention provides a method of producing a rice plant and/or seed for application of a glyphosate-containing herbicide by glyphosate-resistant strain comprising the strain 17053 and comprising SEQ ID N: 14seq ι〇n〇: Self-crossing by plants 'There are seeds produced, seeded to produce progeny plants, progeny plants treated with glyphosate and progeny plants resistant to glyphosate are selected. The above and other aspects of the invention will become more apparent from the following embodiments. "Sequence of sequence SEQ ID NO: 1-- indicates the sequence of twenty nucleotides of the 5' junction sequence between the rice gene DNA and the integrated expression card maker. The nucleotide sequence 147377.doc 201040274 corresponds At positions 565 to 584 of SEQ ID NO: 3 ([C], see Figure 1) and corresponding to positions 565 to 584 of SEQ ID NO: 6. SEQ ID NO: 2-- indicates integrated performance cassettes and rice a sequence of twenty nucleotides of the 3' junction sequence between the genomic DNA. The nucleotide sequence corresponds to positions 626 to 645 of SEQ ID NO: 4 ([D], see Figure 1), and SEQ ID The reverse complement of NO: 2 corresponds to position 3707 to 3726 of SEQ ID NO: 6. SEQ ID NO: 3 - the inserted DNA flanked by line 17053 until X comprises the 5' sequence of the gene transfer DNA region. ID NO: 3 nucleotide positions 5 65 to 584 correspond to nucleotide positions 1 to 20 of SEQ ID NO: 1 and nucleotide positions 575 to 584 of SEQ ID NO: 3 correspond to SEQ ID NO: 5 Nucleotide position 1 to 10. SEQ ID NO: 4-- The inserted DNA flanking the line 17053 up to and including the 3' sequence of the transgene DNA insertion region. SEQ ID NO: 4 nucleotide position 626 to 645 corresponding At nucleotide positions 1 to 20 of SEQ ID NO: 2, and the reverse complementary strand of nucleotide positions 636 to 645 of SEQ ID NO: 4 corresponds to nucleotide positions 3 133 to 3142 of SEQ ID NO: SEQ ID NO: 5-- The sequence of the integrated expression cassette that confers tolerance to glyphosate herbicide. SEQ ID NO: 5 corresponds to nucleotide positions 575 to 3716 of SEQ ID NO: 6. SEQ ID NO: 6-- indicates the 5' sequence (SEQ ID NO. 3) of the inserted DNA flanked by line 17053, the sequence of the integrated expression cassette (SEQ ID NO: 5), and is flanked by line 17053. The nucleotide sequence of the contig of the 3' sequence of the inserted DNA (reverse complement of SEQ ID NO: 4) is 147377.doc 201040274. SEQ ID NO: 7 - used to identify line 17053 Primer SQ4194. The primer SQ4194 is complementary to the region of the gene region flanked by the inserted 5' region of the display cardiner, near the right edge of the insertion of the transgenic DNA. Using the ancestors of the primers SQ4194 and SQ4191 (SEQ ID NO: 8) The amplicon indicates the presence of line 17053. SEQ ID NO: 8-- is used to identify primer SQ4191 of line 17053. The primer SQ4191 is complementary to the inserted 5' region of the expression cassette near the insertion edge of the transgene. The amplicon generated using the combination of primer SQ4194 (SEQ ID NO: 7) and SQ4191 indicates the presence of line 17053. SEQ ID NO: 9 - probe PB1494 to identify line I7053, which is complementary to 5, a fragment of the ligating sequence. The probe can be connected to a decodable marker such as 6FAMTM. The release of a fluorescent signal in combination with a primer such as SQ4194 and SQ4191 and a probe PB1494 such as the labeled 6FAMTM was used in the TAQMAN® (PE Applied Biosystems, Foster City, CA) assay to identify the presence of line 17053. SEQ ID NO: 10--Indicator SQ1 875 ° used to identify line 17053 The primer SQ 1 875 is complementary to the inserted expression cassette 3, region near the insertion edge of the transgenic DNA. Amplicon generated using a combination of primers SQ1875 and SQ3623 (SEQ ID NO: 11) indicates the presence of line 17〇53. SEQ ID NO: 11 - 623 for identifying the line 17〇53. The primer SQ3623 is complementary to the region of the gene region flanked by the inserted 3' end of the expression cassette, and the insertion of the DNA near the transgenic gene. The amplicon generated using the combination of primer SQ1875 (SEQ ID NO: 1〇) and SQ3623 indicates the presence of line 147377.doc 201040274 17053. SEQ ID NO: 12 - primer SQ1871 used to identify flanking genomic DNA of line 17053. The primer SQ1871 is complementary to the inserted 5' region of the inserted cassette near the insertion edge of the transgenic gene DNA. SEQ ID NO: 13 - primer SQ1869 used to identify flanking genomic DNA of line 17053. The primer SQ1869 is complementary to the inserted expression cassette 5· region near the insertion edge of the transgenic gene DNA.

0 SEQ ID NO: 14 - primer SQ1880 used to identify flanking genomic DNA of line 17053. The primer SQ1880 is complementary to the inserted expression cassette 3 region near the insertion edge of the transgenic gene DNA. SEQ ID NO: 1 5--Introduction SQ3626 for identifying flanking genomic DNA of line 17053. The primer SQ3626 is complementary to the region of the gene region flanked by the inserted E5' end of the k-card, near the insertion edge of the transgenic DNA. [Embodiment] The following definitions and methods are provided to better define the present invention and to guide those skilled in the art in the practice of the present invention. Unless otherwise stated, they are understood by those of ordinary skill in the art in light of the The term "comprising" as used herein means "including but not limited to". The present invention provides a line 1 7053 which exhibits a commercially acceptable tolerance to the application of a glyphosate herbicide and a genetically modified rice plant comprising line 17053. This line contains a single-insertion of the gene-transferred DNA into the chromosome/gene of the rice germplasm. Plants comprising lines can be produced by the following steps: (i) transformation of plant cells with a nucleic acid construct comprising a gene of interest, 147377.doc 201040274 (Π) resulting from insertion of a transgenic gene into a plant genome Plant population regeneration, and (iii) selection of specific plants characterized by insertion of the transgenic gene into a specific location of the plant genome. The term "line" refers to a gene-specific insertion that is of interest in a particular location in a genome. A plant comprising a line may be an original transformant comprising a transgenic gene inserted into a specific location of a plant genome. A plant comprising a line may also be a progeny of a transformant comprising a transgenic gene inserted into a specific location of a plant genome. The progeny can be produced by sexual outcrossing between the transformant or its progeny and another plant. The other plant may be a genetically transgenic plant comprising the same or different transgenic genes and/or a non-genetically transgenic plant such as from a different species. Even after repeated backcrossing (back_cr〇ssing) with the recurrent parent, the inserted DNA and the flanking DNA of the transformed parent are still present in the same genetic locus of the progeny. The term "us" in the context of this article means rice (all less and includes all plant species available for rice breeding, including wild-type rice varieties and plants belonging to the rice genus that allow inter-species breeding. "Line" also refers to a DNA molecule from the original transformant comprising the inserted 2Dna and the flanking rice gene dna directly adjacent to either side of the inserted DNA. The DNA molecule is transferred to the rice by inserting the gene into the DNA. The behavior in a plant genome, that is, by a transformation behavior. The DNA molecule thus comprises a nucleotide sequence unique to the rice plant genome in which the gene is inserted and into which the gene has been inserted, wherein the core The nucleotide sequence contains the sequence of the specific region of the genomic DNA and the sequence of the inserted gene-transferred dNA. The DNA inserted in the rice line 17053 is aligned with the surrounding rice plant base 147377.doc •10-201040274 due to the arrangement of the body DNA It is unique and unique to the rice line m53. The DNA molecule is also a component of the rice chromosome of the plant containing the line i m3 'and thus is statie in the plant t and can be passed to the plant. 卞. Line 1 7 0 5 3 confers tolerance to glyphosate weeding swords applied to rice plants. "Glyphosate" means N-phosphinylmethyl-glycine and its salts. Glycosylglycine is a herbicide that is active against a wide range of plant species. When applied to the surface of plants, glyphosate moves within plants by systemic action. Glyphosate has a pathway to inhibit the shikimate pathway. Phytotoxicity, the oxalic acid pathway provides a precursor for the synthesis of aromatic amino acids. The term "recombinant" as used herein refers to DNA and/or protein that is not normally found in nature and thus is formed by human intervention. / or have a 'body form. This human intervention can produce recombinant DNA molecules and / or recombinant plants. As used herein, "recombinant DNA molecule" is a combination of DNA molecules that contain results that are not naturally present but are human intervention. a Q DNA molecule 'eg, a DNA molecule comprising a combination of at least two DNA molecules that are heterologous to each other; and/or synthetically and comprising a polynucleotide sequence that differs from the polynucleotide sequence normally found in nature. a dna molecule; and/or a DNA molecule comprising a transgenic gene artificially incorporated into the genomic DNA of the host cell and a DNA flanked by the host cell genome. Examples of the recombinant DNA molecule are the insertion of the transgenic gene as described herein. a DNA molecule produced in the rice genome, which may ultimately cause the recombinant rnA and/or protein molecule to be expressed in the organism. As used herein, "recombinant plant" is a plant that is not normally found in nature. For human intervention, 147377.doc -11- 201040274 contains and contains a transgenic gene and/or a heterologous DNA molecule incorporated into its genome. Because of this genomic alteration, 'recombinant plants are clearly different from related wild-type plants. An example of a recombinant plant is a rice plant comprising the line n〇53 as described herein. The term "transgenic gene" as used herein refers to a polynucleotide molecule that is artificially incorporated into the genome of a host cell. The transgene can be a heterologous gene relative to a host cell. The term "genetically transferred plant" refers to a plant comprising the transgenic gene. The term "heterologous" as used herein means that a combination of a first molecule and a second molecule is not normally found in nature. For example, a molecule can be derived from a first species and inserted into a genome of a second species. The molecule is therefore heterologous to the host and is artificially incorporated into the host cell genome. The term "dissociation" as used herein refers to a single dna molecule produced by fusing a first DNA molecule with a second DNA molecule, wherein the first or second DNA molecule is generally not found to be in a conformational form ( That is, it is integrated with the other). After the DNA molecule, the Helmet A is a new DN Α molecule that is usually not found in nature.桊Invention provides DNA molecules, μ % qq «also uses sigma + and the terms "DNA", "eighth" first NA knife, "polynucleotide molecule" derived from the genome or synthesis of the double clock λ8 The chain DNA molecule, which is the polymer of the deoxyribonucleic acid base or the upstream of the polynucleotide, is read to the end of the 3, (-) end. As used herein, the term "DNA sequence sequence" or "polynucleotide sequence at 4t" refers to the nucleotide/column of a DNA molecule. The terminology used in this article is stipulated in Title 3 of the Federal Regulations. 147377.doc -12- 201040274 Section 1.822 is required and displayed in the tables of WIPO Standard ST.25 (1998), Appendix 2, Tables 1 and 3. . The present invention is disclosed with reference to SEQ ID NOS: 1-6 with reference to SEQ ID NO: 1-6, with reference to only one of the two nucleotide sequence strands present at the position of the gene-transferred DNA inserted in the gene of the gene-transgenic line 17053. Thus, by implication and derivation, a complementary sequence (also referred to in the art as a complete complementary sequence or a reverse complementary sequence) is within the scope of the invention and is therefore intended to fall within the scope of the claimed subject matter. The nucleotide sequence corresponding to the entire nucleotide sequence of the inserted gene-transferred DNA and the substantial segment of the rice genomic DNA flanked at either end of the inserted gene-transforming DNA is herein SEQ ID NO : 6 available. The inserted gene-transferred DNA provided in SEQ ID NO: 5 is a segment in this. The nucleotide sequence of the genomic DNA which is ligated to the 5' end of the inserted gene transfer DNA (SEQ ID NO: 5) by a phosphodiester bond is SEQ ID NO: 3 Show. The nucleotide sequence of the genomic DNA which is ligated to the 3' end of the inserted gene transfer DNA (SEQ ID NO: 5) by a phosphodiester bond is SEQ ID NO: 4 Show. Line 1 7053 additionally comprises two polynucleotide sequences, one covering the 5' position and one covering the 31 position, wherein the gene-transforming DNA is inserted into the somatic DNA, which is referred to herein as the junction sequence. The "splicing sequence" or "joining region" refers to a DNA sequence and/or a corresponding DNA molecule covering the inserted gene-transferred DNA and the adjacent flanking DNA. The ligated sequence is arbitrarily represented by two sequences of 20 nucleotides each and is provided as SEQ ID NO: 1 and SEQ ID NO: 2, each of which represents 10 nucleosides of directly adjacent flanking DNA. The acid is attached to 10 nucleotides of the inserted DNA. The nucleosides 147377.doc -13- 201040274 acid are linked by a phosphodiester bond. In rice, SEQ ID N〇: 1& SEQ ID NO: 2 is not naturally occurring in the genome and is unique and unique to the gene transfer line 17〇53, and is derived from any nucleotide sequence derived from a rice plant. Recognition of at least about 、, at least about 丨3, or at least about 15 contiguous nucleotides for each of the sequences, or the like, is conclusive evidence that the DNA is obtained from the rice line 17〇53, And the presence of DNA from the rice line i 7〇53 in the sample was identified. SEQ ID NO: 1 is the sequence of 20 nucleotides covering the junction between the rice gene DNA and the inserted DNA. Seq id NO: 2 is a sequence of 2 nucleotides covering the junction between the rice gene DNA and the 3' end of the inserted DNA. The invention thus provides a DNA molecule comprising at least the nucleotide sequence set forth in SEq ID NO: 1 and/or SEQ ID NO: 2. Any segment derived from the DNA of the genetically modified rice line i 7 〇5 3 comprising at least about 11, at least about 3 or at least about 5 contiguous nucleotides of SEQ ID NO: 1 is within the scope of the invention The scope. Any segment of DNA derived from the gene-transgenic rice line 17053 that is sufficient to include at least about 11, at least about 13, or at least about 15 contiguous nucleotides of SEQ ID N: 2 is within the scope of the invention. Furthermore, any polynucleotide comprising a complementary sequence to any of the sequences described in this paragraph is within the scope of the invention. Figure i illustrates the physical arrangement of SEQ ID n 〇, 1-5 relative to SEQ ID NO: 6 from 5, to 3, arranged. The invention also provides at least 80%, 85°/. , 90%, 95%, 97%, 98% or 99% of the nucleic acid molecule of SEQ ID NO: 6. The invention provides exemplary DNA molecules that can be used as primers or probes to identify the presence of DNA derived from line 17053 in a sample. Such primers or probes are specific for the target nucleic acid sequence and are thus suitable for use in the method of the invention described herein by 147377.doc 14-201040274 to identify rice line 17053. "Introduction j is typically a highly purified, isolated polynucleotide designed for specific adhesion or hybridization methods involving thermal amplification. It can be used in thermal amplification (such as polymerase chain reaction (PCR)). Template DNA (such as a sample of rice genomic DNA) and a pair of primers to generate an amplicon, wherein the amplicon from the reaction will have a sequence corresponding to the template DNA that is hybridized between the primers to the two points of the template. DNA sequence. As used herein, "amplifying q" is a piece of DNA or a piece of DNA that has been synthesized using amplification techniques. In one embodiment of the invention, the amplicon of line 17053 is identified to comprise a sequence that is not found in the rice genome. The amplicon of the present invention comprises at least about contiguous nucleotides of Seq id NO: 1, SEQ ID NO: 2 and/or its complement, at least about 13 contiguous nucleotides, or at least about 丨At least about 3 or to - about 15 contiguous nucleotides. The primer is usually designed to hybridize with the complementary target 1)1^8-strand to form a hybrid between the primer and the target, and the primer is used for the polymerase to use the target DNA strand as a template to start the extension of the primer (also to other nucleotides) A recognition site that is polymerized into an extended polynucleotide molecule). When a primer pair is used in the present invention, it is meant that in a thermal amplification reaction or other conventional nucleic acid amplification method, two primers that bind to opposite strands of a double-stranded nucleotide segment are used to achieve linear amplification of the primer pair. The purpose of the polynucleotide segment between the target positions of the individual elements is combined. SEQ IDs 7-8 and 10-11 are provided as exemplary molecules suitable for use as primers. A primer pair as provided in SEQ ID NO: 7 and SEQ ID NO: 8 is provided as a first DNA molecule and a second DNA molecule different from the first DNA molecule, and each of the two molecules has SEQ ID NO: 3, SEQ ID NO: 5 or SEQ ID NO: 6 147377.doc -15- 201040274 A sufficient length of contiguous nuclei: g: acid or its complementary sequence to serve as a thermal expansion of the dna lead + 'f and the (4) rice line 17G53 template face Increasing the reaction to generate at least about (10) contiguous nucleotides comprising SEQ ID NO: 丨, at least about! 3 contiguous nucleotides, or at least about (1) solid, at least (four) or at least about 15 contiguous (tetra) acids. child. An exemplary DNA molecule pair is provided, ie, SEQ ID NO: 1 〇 and SEQ ID 〇: u, as a first DNA molecule and a second molecule different from the first DNA molecule, and each of the two molecules has SEQ m NO: 4, SEQ A sufficient length of contiguous nucleotide acid of ID Ν0·5 or SEQ ID N〇6 or a complementary sequence thereof to serve as a DNA primer for use together with a ruthenium amplification reaction derived from a template DNA derived from rice line 17053 At least about four (4) contiguous nucleotides, at least about 13 contiguous nucleotides, or at least about (1) solid, at least (four) or at least about 15 contiguous nucleotides of SEQ m. A sputum needle is an isolated nucleic acid that is complementary to a strand of a target nucleic acid. The probe of the present invention not only includes deoxyribonucleic acid or ribonucleic acid which specifically binds to a target ship sequence, but also includes a specific binding target dna sequence, guanamine and other probe materials, and the combination (4) can be used for the clock Determining, identifying, determining, or determining the presence of the target DNA sequence in a particular sample. The probe can be linked to a well-testable conventional label or reporter molecule (e.g., a radioisotope, a ligand, a chemiluminescent agent, or an enzyme). In one embodiment of the invention, the probe of the 'Qingding Line 17G53 comprises a sequence that is not naturally found in the rice genome. SEQ ID NO: 9 is provided as an exemplary dna molecule suitable for use as a probe. The probes and primers of the present invention may have a complete sequence with the target sequence - 147377.doc 201040274, and it is possible to design a primer which is different from the target sequence and retains the ability to preferentially hybridize with the target b sequence by a conventional method. Probe. In order for a nucleic acid molecule to act as a primer or probe, it only needs to be sufficiently complementary in sequence to be able to form a stable double-stranded structure at the particular solvent and salt concentration employed. Any of the known nucleic acid hybridization or amplification methods can be used to identify the presence of the transgenic DNA of the rice line 17053 in the sample. Probes and primers are typically at least about II nucleotides, at least about 18 nucleotides, at least about 24 nucleotides, or at least about 3 nucleosides or more. The probes and primers specifically hybridize to the target DNA sequence under stringent hybridization conditions. The strict conditions are known by Sambr〇〇k et al., I., and the gambling of others, in Nuchic HybHdizati〇n, mouth/VacOca/ 洲洲, iRL Press, Washingt〇n, ((10)5). As used herein, two nucleic acid molecules are said to be capable of specifically hybridizing to each other if they are capable of forming an anti-para-ar double-stranded nucleic acid structure. If it exhibits complete complementarity, one nucleic acid molecule is referred to as the "complementary sequence" of the other nucleic acid molecule. As used herein, when each nucleotide of one molecule is complementary to the nucleotide of the other, the two molecules are said to exhibit 70 full complementarity. The two molecules are said to be "minimally complementary" if they are sufficiently stable to hybridize to each other to adhere to each other under at least the "low stringency" conditions. Similarly, two molecules are said to be "complementary" if they are sufficiently stable to hybridize to each other under known "high stringency" conditions. Thus deviations from complete complementarity may be allowed as long as such deviation does not completely impede the ability of the molecule to form a double-stranded structure. The term "isolated" as used herein refers to at least partially separating a molecule from other molecules that it normally associates with in its original or natural state. 147377.doc -17· 201040274 In one embodiment, the term "isolated" refers to a DNA molecule that is at least partially separated from a nucleic acid that is normally flanked by the DNA molecule in its original or native state, such that, for example, due to recombinant techniques, DN A molecule is fused to a regulatory or coding sequence that is not normally associated therewith and is considered isolated herein. Even if the molecules are integrated into the host cell chromosome or are present in a nucleic acid solution containing other DNA molecules, they are considered to be isolated. Many methods well known to those skilled in the art can be used to isolate and manipulate DNA molecules or fragments thereof that are not disclosed in the present invention. For example, PCR (polymerase bond reaction) techniques can be used to amplify specific starting DNA molecules and/or to generate variants of the original molecule. DNA molecules or fragments thereof can also be obtained by other techniques, such as direct synthesis of fragments by chemical methods, which are typically carried out by using an automated oligonucleotide synthesizer. The DNA molecules and corresponding nucleotide sequences provided herein are therefore particularly suitable for identifying rice lines 17053, selecting plant varieties or hybrids comprising rice lines 17〇53, detecting the presence of the rice line 17〇532 DNA from the sample and Samples were monitored for the presence and/or absence of rice line 17053 or plants and plant parts containing rice line 17053. The present invention provides rice plants, progeny, seeds, plant cells, plant knives (such as t private, ovule, inflammation, flower, root or stem tissue, fibers and leaves) and commodities. Such plants, progeny, seeds, plant cells, plant parts and lysates have detectable amounts of the polynucleotide of the invention, ie such as having at least one such as SEQ ID NO: 1 and SEQ ID Ν 0: 2 The sequence of the provided polynucleosides. The plants, progeny, seeds, plant cells and plant parts of the invention may also contain one or more other transgenic genes. The transgene may be any nucleotide sequence encoding a protein or RNA molecule that confers the desired properties to 147377.doc -18-201040274, including but not limited to enhanced insect resistance, increased water use Efficiency, increased yield performance, enhanced drought resistance, enhanced seed quality, improved nutritional quality, and/or enhanced herbicide tolerance, where the desired knowledge is relative to rice lacking the other transgenic genes Plant measurement. The present invention provides rice plants, progeny, seeds, plant cells, and plant parts comprising lines 17053 (such as powders, ovules, pods, flowers, roots or stem tissues and leaves derived from genetically modified rice plants). Representative seed samples containing lines 17〇53 have been deposited under the Budapest Treaty for the operation of the present invention. The storage facility selected for receiving storage is the American Type Culture Collection (AT CC) at 10801 University Boulevard, Manassas, Vlrgmia USA, USA. Area code 20U0. The ATCC storage designated line 17053 seed registration number is PTA-9843. The present invention provides a microorganism comprising a DNA molecule having SEQ ID N: SEQ ID NO: 2 present in its genome. An example of such a microorganism is a gene transfer plant cell. Microorganisms, such as plant cells of the invention, are suitable for use in a variety of industrial applications including, but not limited to: (1) as a research tool for scientific inquiry or industrial research; (ii) for culture to produce endogenous or recombinant carbon water Compounds, lipids, nucleic acids or protein products or small molecules, which can then be used for scientific research or as industrial products; and (丨(1) can be used in modern plant tissue culture techniques to produce genetically transgenic plants or plant tissue cultures, which can be used after Ik Agricultural research or production. Preparation and use of microorganisms (such as gene transfer plant cells) using modern microbiology techniques and human stem 147377.doc -19- 201040274 pre-production of unique artificial microorganisms. In this process, recombinant DNA is inserted into plants. A gene transfer plant cell that is separate and unique from the naturally occurring plant cell is formed in the cell genome. The gene transfer plant cell can then be cultured using modern microbiological techniques like bacteria and yeast cells and can exist in an undifferentiated, single-cell state. The genetic composition and phenotype of new plant cells is to integrate heterologous DNA into fine Technical effects formed in the genome. Another aspect of the invention is the method of using the microorganism of the invention. The method of using the microorganism of the invention (such as a gene transfer plant cell) comprises (1) by integrating the recombinant DNA into the cell genome And subsequent use of the cells to obtain other cells having the same heterologous DNA to produce gene-transgenic cells; (n) methods for culturing cells containing recombinant DNA using modern microbiological techniques; (iii) production from cultured cells And methods for purifying endogenous or recombinant charcoal water compounds, lipids, nucleic acids or protein products; and (iv) using genetic engineering tissue culture techniques to genetically transfer plant cells to produce gene transfer plants or gene to lean plant tissue cultures The plant of the present invention can deliver strain DNA (including a transgene) to a progeny. As used herein, "progeny" includes any strain comprising DNA derived from an ancestral plant and/or comprising at least one such as SEQ. Plant NO, Plant, Seed, Plant Cell and/or Reproductive Plant Department of ID NO: 1 and SEQ ID NO: 2 Plants, progeny and seeds may be homozygous or heterozygous for the transgenic genes. Progeny may be grown from seeds produced by rice plants containing lines 17〇53 and/or from rice plants containing lines 17〇5 3 Seeds produced by fertilized fertilized plants. 147377.doc -20- 201040274 Progeny plants can be self-pollinated (self_p〇llinated, also known as "selfmg") to produce pure breeding lines. A plant (that is, a homozygous plant of a transgenic gene). Selfing of appropriate progeny can result in a homozygous plant of two exogenous genes. Alternatively, the progeny plant can be outcrossed, for example, with another Related plant breeding produces variants or parental seeds or plants. Another unrelated plant can be a gene transfer plant or a non-gene transfer plant. Thus, a variant or a hybrid seed or plant of the invention may be derived from a first parent lacking the unique line of the rice line 17〇53 and having a distinct dna, and a second parent comprising the rice line 17〇53, resulting in a rice line containing 17053 A unique and unique hybrid of DNA. Each parent may be a hybrid or an inbred/variant, as long as the hybrid or breeding produces the plant or seed of the invention, ie the seed has at least one allele. Contains the unique and unique rice line 17053 DNA comprising SEQ ID NO: 1 and SEQ ID NO: 2. Thus, two different gene transfer plants can be mated to produce hybrid progeny containing two independent isolates plus a foreign gene. ◎ For example, a glyphosate-tolerant rice comprising line 1 7053 can be crossed with other genetically transgenic rice plants to produce plants having the characteristics of two gene-transforming parents. An example of this is that a glyphosate-tolerant rice comprising line i 7〇53 is crossed with a rice plant having one or more other characteristics to produce a progeny plant or seed that is tolerant to glyphosate and has eight or more other characteristics. . For example, a rice plant showing phosphinothricin or giuf〇_sinate herbicide tolerance can be crossed with a rice plant comprising line 17〇53 to produce glyphosate-tolerant and glufosinate-resistant or Progeny plants or seeds of glufosinate herbicides Thus, plants and seeds for use in the methods disclosed herein may also contain 147377.doc 201040274 - or a plurality of other transgenic genes. The transgenic gene may have a property of a protein 皙 red ρ, a, a flat code to confer a nucleotide sequence to the shell or RNA molecule, including but not limited to aphid μ Sexuality, increased water efficiency, yield performance, i« strong eve private 3 1 double early, increased θ strong drought resistance, enhanced seed quality, quality, & good-force tolerance and / or enhanced weeding : 2 battalions of the required materials are lacking in material and he is transferred to the raw quantity. Soil M <Non-plant 踯 gene transfer plants have been shown to have tolerance to them and herbicides which can be applied by the present method include (but are not limited to): licorice, turf scale, continuum mm (sulfonylurea) ^ ( Imidazolinone) . ^ ^ (-ynu), thatch (delapon), cyclohexanedione, lin = enzyme (pr〇toporphyri〇n〇gen 〇xidase) inhibitor and isoxasflutole herbicide . Polynucleotide molecules encoding proteins associated with herbicide tolerance are known in the art and include, but are not limited to, polynucleotide molecules encoding the following: glyphosate-tolerant 5-enol Acetone oxalic acid-3-phosphate synthase (EPSPS) (see, for example, U.S. Patent Nos. 'a? 〇6, 5, 633, 435, 6, 040, 497, 5, 〇 94, 945, 5, 804, 425, 6, 248, 876, 7,183,110, No. 39,247); glyphosate oxidoreductase (G0X) (see, e.g., U.S. Patent No. 5,776,760) · glyphosate-η-acetyltransferase (gAT); herbicide resistant The type of acetaminophen lactate synthase (ALS, also known as acetamidine hydroxyacid synthase (AHAS)) is used by the scutellaria sulphate, the sulphate, the ketone, the ketone Pyrimidine ester, sulfonamide carbonyl triazolinone and/or heteroaryl ether; herbicide tolerant acetamyl coenzyme A carboxylase (ACCase) or R-2,4-II 147377.doc -22- 201040274 Chlorophenoxypropionic acid dioxygenase (rdpA) to withstand aryloxyphenoxypropionate (AOPP) (such as haloxyfop, quizalofop) , dichlorofop and diclofop); such as 2,4-D dioxygenase (tfdA), R-2,4-dichlorophenoxypropionic acid dioxygenase (rdpA) , a deoxygenated protein of aryloxyalkanoate dioxygenase (AAD) and/or S-2,4-dichloropropionic acid dioxygenase (sdpA), used to tolerate synthetic auxin Herbicide; bromoxynil nitrilase (Bxn) to tolerate bromoxynil (see, e.g., U.S. Patent No. 4,810,648); phytoene desaturase (crtl) to withstand gas Norflurazon; bialaphos resistance (bar) or glufosinate acetyltransferase (PAT) protein (see, for example, U.S. Patent Nos. 5,646, 〇24 and 5,276,268) for resistance Containing glufosinate and biloba; and using it for triketones (mezotrione, tembotrione, topromezone, iso-evil. sitting) weeding Protein, such as tolerant 4-hydroxyphenylpyruvate dioxygenase (HPPD), detoxified cytochrome P450, or HPPD pathway accessory, such as G. globulin g7o0zy Brwb) HPP oxidase (HPPO) and Pseudomonas acidovorans A-}l'?A 1-Chemase (HPAH) and NADH oxidoreductase (HPAC). Polynucleotide molecules encoding other proteins suitable for use in gene transfer plants are known in the art and include, but are not limited to, polynucleotide molecules encoding the following: E. coli (£. cW) cspA (PCT) Publication No. WO 2005/0333 No. 18); Bacillus subtilis (anti-wZ^/WcspBCPCT Publication No. WO 2005/0333 18); corn (Zm transporter (US 147377.doc -23-201040274 Publication No. 20040434888) , No. 20070011783, No. 20070294782); corn nfb2 (a.k_a hap3) (US Publication No. 20050022266 and No. 20080104730, PCT Publication No. WO 2008/002480); cotton csp (csp-like) ( US Application No. 1 1/980,758, U.S. Publication No. 20050097640, PCT Publication No. WO 2005/0333 18); Wheat Csp (US Patent No. 7,214,786, U.S. Publication No. 20050097640); G1988 (USA) Application No. 9/474, 435, pp. No. 04/031, 349, filed on Jun. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. No. And CGPG 2117 (United States Publication No. 20080090998. Other polynucleotide molecules encoding proteins suitable for gene transfer plants are known in the art and include pest control (such as nematicides, fungicides, and insecticides). The insecticides include, but are not limited to, Bt toxins and/or pathogenic genus, photosynthetic genus (P/zoior/mfei/M·?), Bacillus (such as Bacillus licheniformis) (10,000/aierosporow·?)), iSerraiia, Klebsiella (Ar/ehz'W/fl), Erwinia and similar genus toxins. Bt toxins include ( But not limited to) VIP toxin, Cryl toxin, Cry2 toxin, Cry3 toxin, Cry4 toxin, Cry5 toxin, Cry7 toxin, Cry8 toxin, Cry9 toxin and binary insecticidal toxin (such as those derived from Bt) and similar toxins Like vegetative propagation, it also covers backcrossing with parent plants and outcrossing with non-genetically transformed plants. Descriptions of other breeding methods commonly used for @& and crops can be found in several references 147377 .doc -24· 201040274, Fehr as in Breeding Methods f〇r Cultivar Development, Wilcox J. ed, American s〇ciety 〇fAgr〇n〇my, Madison WI (1987) in. The present invention provides a plant part derived from a rice plant comprising line 17053. As used herein, "plant part" means any part of a plant which is composed of a material derived from a rice plant comprising line 17〇53. Plant parts include, but are not limited to, pollen, ovules, pods, flowers, roots or stem tissue, fibers and leaves. The plant parts can be viable, non-viable, regenerable and/or non-renewable. The present invention provides a product derived from a rice plant comprising the line 17〇53. As used herein, "commodity" means derived from a contained line 1 . <(4) a product or product of a material of a rice plant, seed, plant cell or plant part. Goods can be sold to consumers and can survive or survive. Commodity that does not survive includes (but is not limited to) seeds and weights that cannot survive, seeds, seed parts and plant parts that are processed; dehydrated plants €), tissues, cold tissue and processed plant tissues; And/or animal feed for aquatic animals, 〉, 、,meai, flour, shards, sorghum, fiber, milk, cheese, paper, cream, wine and any other human food Seeds and plant parts, and biomass and fuel production. Commodities that can survive include, but are not limited to, seeds and plant cells. The package 3 ° ° 17053 meters can therefore be used to make any quotient that is usually obtained by seeking. Any such product derived from a plant comprising line 17053 can contain a unique and unique amount corresponding to the ν detectable amount of 米7〇53 and specifically contain detectable amounts of SEQ IDN0 : 丨I47377.doc -25- 201040274 or a polynucleotide of at least 15 contiguous nucleotides of SEQ ID NO: 2. Any standard detection method using a polynucleotide molecule can be used, including the detection methods disclosed herein. If any detectable amount of SEQ(1)N〇: 1 or SEQ ID NO: 2 ' is present in the commodity, the article falls within the scope of the invention. Plants, progeny, seeds, plant cells, plant parts (such as powders, ovules, pods, flowers, roots or stem tissues and leaves) and commodities of the invention are therefore particularly suitable for planting plants to produce seeds comprising lines 17〇53 And/or plant parts are used for agricultural purposes, producing progeny containing line i7〇53 for plant species and research purposes, using microbiological techniques for industrial and research applications' and selling to consumers. The present invention provides methods of using glyphosate herbicides and plants comprising lines 17〇53 to control weeds and methods of producing plants. Providing a method of controlling weeds in a field comprising planting a variety or hybrid plant comprising lines 17〇53 in the field and using a weeding dose of grass (4) in Xiaotian 16 for (iv) field weeds but not plants containing line 17G53 . The application of the herbicide herbicide can be carried out before emergence (i.e., at any time after planting the seed containing the line m53 and before emergence of the plant containing the line 17053), or after emergence (i.e., emergence of the plant containing the line 17053) After any time). Also, for another method of controlling field weeds, the method comprises applying an effective agent 2 diglyphosate herbicide to control weeds in the field and then planting rice of 170353 in the field. The application of the glyphosate herbicide should be carried out before planting, ie before seeding of the line containing m53, and can be carried out before any planting, including (but not limited to) about 14 days before planting until before planting. The herbicidal effective dose of glyphosate used in the field for about 7 days will be between about 〇5 lbs/acre of glyphosate to about 45 lbs/acre in the growing season by Fan I47377.doc -26- 201040274. Glyphosate composition. The grass can be applied multiple times during the growing season, for example, two applications (such as pre-plant application and post-emergence application or pre-emergence application and post-emergence application) or three applications (such as pre-plant application, pre-emergence application and post-emergence application). Application). • A method of producing a rice plant comprising a DNA sequence unique to the gene-transgenic line of the present invention and having a unique DNA sequence and tolerance to a herbicide. The genetically transgenic plants used in these methods may be homozygous or heterozygous for the transgenic gene. The progeny plants produced by such methods may be varieties or hybrid plants; the rice-containing lines produced by the seeds produced by the plants comprising line 17053 and/or by the plants fertilized with the pollen of the rice plants containing line 17053 The seed of 17053 grows; and it can be a homozygous junction of the transgenic gene or a . The progeny plants can then be self-pollinated to produce plants of the pure breeding line (ie, plants that are homozygous for the transgenic genes), or can be cross-extracted, for example, with another species of plant, which produces variants or hybrids. Seed or plant. Plants comprising a strain of a molecule comprising a sequence of SEQ m N〇:丄 and 〇EQ ID NO. 2 can be sexually crossed with another rice plant and thereby produce a seed, which is subsequently grown A progeny plant is produced to produce a rice plant that is applied by a glyphosate herbicide. The progeny plants can then be treated with a glyphosate herbicide to select progeny plants that are tolerant to the herbicide herbicide. Alternatively, the progeny plants can be analyzed using an identification method to select progeny plants containing line 17053 DNA. The other plant for hybridization may or may not tolerate the glyphosate herbicide and may be a genetically transgenic plant or may not be a genetically transformed plant. The progeny plants and/or seeds produced may be varieties or hybrid seeds. 147377.doc -27- 201040274 In the case of sputum, the step of sexually crossing a plant with another plant (ie, cross-pollination) can be achieved or facilitated by human intervention, for example by collecting the pollen of the plant by hand. Exposing the pollen to the style of the second plant or removing or destroying or stabbing the stamens or anthers of the plant by hand and/or behavior (for example, by emasculation or by applying a chemical gametocide) Self-pollination is obstructed and cross-pollination must be carried out to achieve fertilization; artificial pollination is carried out by placing seeds at the "direct pollination" position (9) by placing beehives in orchards or in the fields or by trapping plants with pollinating insects ) by manually opening or removing parts of the flower to place or contact the external pollen on the style or stigma (for example, rice naturally prevents or prevents cross-pollination of flowers without human intervention) Naturally self-pollination; by selective placement of plants (eg, intentionally planting plants near pollination); and/or by early application of chemical agents to promote flowering or to promote receptivity (column for ^ Powder acceptance). Plants can be produced by self-crossing a plant comprising line 17053 and comprising a polynuclear (tetra) molecule comprising the sequence of SEq ID N〇: i and SEQ ID N〇: 2 and thereby producing a seed, which subsequently grows into a progeny plant Rice plants applied by glyphosate herbicides. The progeny plants can then be treated with a glyphosate herbicide to select progeny plants that are tolerant to glyphosate herbicides. Alternatively, the progeny plants can be analyzed using an identification method to select progeny plants containing the line DNA. In practicing this method, the step of sexual hybridization (ie, self-pollination or self-crossing) of a plant with itself can be achieved or facilitated by human intervention, for example, by collecting the pollen of the plant by hand and bringing the pollen into contact with 147377. Doc -28- 201040274 The style or stigma of the same plant and subsequent prevention of re-fertilization of the plant as appropriate. · Removal, destruction or obscuration of stamens or anthers of other nearby plants by hand and/or behavior (eg by emasculation or By applying chemical gametes • Qi!) to prevent natural cross-pollination from being polluted and self-pollination must be achieved • Fine; artificially placed pollinators at the location of “direct pollination” (eg by pollinating insects) Trapping only one plant); self-pollination by artificial manipulation of flowers or parts thereof; by selective placement of plants (eg, intentionally planting plants outside the pollination); and/or early flowering by application of chemical agents Or promote acceptance (the stigma is acceptable for pollen). The progeny rice (4) and seeds covered by the methods and produced by using the methods will be different from other rice plants, for example due to the progeny rice plants and seeds: reconstituted and thus formed by human intervention; a herbicide herbicide; comprising at least one allele consisting of the DNA of the gene of the present invention; and/or comprising a detectable amount of a group selected from the group consisting of seq(1) NO. 1 and SEQ ID NO: Core: g: acid sequence. The seed may be selected from the individual progeny plants' and as long as the seed comprises seq id N〇: ^ and SEQ ID NO: 2, it is within the scope of the invention. In the practice of the present invention, two different genetically transgenic plants can be crossed to produce hybrid progeny containing two independently isolated heterologous genes. Selfing of appropriate progeny can result in plants that are homozygous for both genes. Like vegetative reproduction, it also covers backcrossing with parent plants and outcrossing with non-genetically transformed plants. Descriptions of other methods commonly used for different characteristics and crops can be found in several references, such as Fehr in Breeding Methods for Cultivar

DeVel〇pment, edited by WileGX L, Αη^^η—My Ag and 〇; 147377.doc -29· 201040274

Madison WI (1987). Plants and seeds for use in the methods disclosed herein may also contain one or more other transgenic genes. The transgenic gene can be any nucleotide sequence encoding a protein or RNA molecule that confers the desired properties, including but not limited to: 3⁄4 strong insect resistance, increased water efficiency, increased yield performance , enhanced drought resistance, enhanced seed quality, improved nutritional quality and/or enhanced herbicide tolerance, the desired characteristics in # are measured relative to rice plants lacking the other transgenic genes. The method according to the invention is therefore particularly suitable for controlling weeds in the field while planting plants in order to produce seeds and/or plant parts comprising line m53 for agricultural or research purposes, selecting progeny comprising lines 17〇53 for plant breeding or research. (d) 'and produce progeny plants and species containing the line 17G53. Plants, progeny, species early, μ & i & \path to plant cells, plant parts (such as powder, ovule, English, flower, root) Or stem tissue and leaf) and commodities may be evaluated for DNA composition, gene expression and/or protein performance. The assessment can be performed by using standard methods such as PCR, Northern blotting, Southern analysis, Western blotting, immunoprecipitation, and ELISA, or by using the detection methods and/or detection kits provided herein. . And provide a method for the presence of materials unique to the rice bran mouth m 5 3 in the test sample. A method comprising detecting the presence of DNA specific to and derived from a rice cell, tissue or plant comprising line 17053. The method provides a template DNA sample to contact a primer pair that is capable of producing an amplicon of strain 17G53 under conditions suitable for thermal amplification, particularly comprising _(1) 147377.doc -30- 201040274 NO. 1 or SEQ ID NO: An amplicon of at least 15 contiguous nucleotides of 2 or a complement thereof. The amplicon is produced from a template DNA molecule derived from rice line 17053 'as long as the template DNA molecule incorporates a unique and unique nucleotide sequence as set forth in SEQ ID NO: 1 and SEQ ID NO: 2. The polymerase' amplicon selected for generating an amplicon may be single-stranded or double-stranded Ο Ο D N A or R N A . The method can be used to detect an amplicon molecule produced in any of the thermal amplification reactions, and to confirm that the nucleotide corresponding to SEQ ID N:: or SEQ ID NO: 2 or its complementary sequence in the amplicon sequence presence. Detecting the nucleotide corresponding to SEQ ID NO: 1 or SEQ ID NO. 2 or its complement in the amplicon, and determining and/or identifying the presence of the DNA specific to the line 17〇53 in the sample and thus Contains the presence of a biological material of line 17〇53. Another method for detecting samples consisting of materials derived from rice plants or rice plant tissues corresponds to SEQ ID n: 3 and π. NO. 4, the presence of DNA molecules. The method consists of: (1) extracting a DNA sample from a rice plant or a group of different rice plants, and (ii) contacting the DNA sample to exhibit at least 15 as shown in SEQ ID NO: 1 or SEq ID N〇:眶 probe molecules adjacent to the nucleus (4), (4) hybridization of the probe and brain sample under stringent hybridization conditions, and subsequent (iv) mapping of the hybridization line between the probe and the target DNA sample. Depending on the case, the detection of the hybridization composition in the 匪 sample identifies the presence of SEQ(1)n〇: 3 or 卿ι〇 NO. Alternatively, there is no hybridization identification sample in the absence of a gene transfer system or 'measure that a particular rice plant contains any of the sequences corresponding to SEQ ID NO: 1 or SEQ ID Ν〇: 2, or both, or complement each other The sequence 'W' the rice plant contains at least one 147377.doc-31 · 201040274 allele corresponding to line 17053. Thus, the presence of the nucleic acid molecules of the present invention can be detected by any well-known nucleic acid detection method such as polymerase chain reaction (PCR) or DNA hybridization using a nucleic acid probe. A strain-specific PCR assay is described, for example, by Taverniners et al. (J. Agric. Food Chem., 53: 3041-3052, 2005), which demonstrates the use of the gene-transforming maize lines Btll, Btl76 and GA21 and for A strain-specific tracking system for the mustard line RT73. In this study, line-specific primers and probes were designed based on the sequence of the genomic/transgenic junctions of each line. Gene-transplanting plant line-specific DNA detection methods are also described in U.S. Patent Nos. 6,893,826, 6,825,400, 6,740,488, 6,733,974, 6,689,880, 6,900,014 and 6,818,807. Provide a DNA detection kit. A kit of the type comprising at least one DNA molecule having a sufficient length contiguous nucleotide similar or complementary to SEQ ID NO: 3, SEQ ID NO: 5 or SEQ ID NO. 6 to serve as a source for specific detection of the sample A DNA primer or probe that is present in the DNA of the gene-transgenic rice line 17053. The DNA molecule detected by the kit contains at least 15 contiguous nucleotides as shown in SEQ ID NO: 1 or a complement thereof. Alternatively, the kit may contain at least one DNA molecule having a sufficient length contiguous nucleotide similar or complementary to SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6, serving as a source for specific detection of biological samples. A DNA primer or probe that is present in the DNA of the gene-transgenic rice line 1 7053. The DNA molecule detected by the kit contains at least 15 contiguous nucleotides as shown in SEQ ID NO: 2 or a complement thereof. 147377.doc -32- 201040274 An alternative kit employing a primer pair that contacts a target DNA sample as described above, followed by amplification sufficient to produce at least 15 contiguous nucleotides comprising SEQ ID NO: 2 or complementary sequences thereof The method of the nucleic acid amplification reaction of the enhancer. Detection of the amplicon and determination of the presence of not less than 15 contiguous nucleotides or their complements of the sequence RSEQ ID NO: 1 or SEQ ID NO: 2 of the amplicon can be determined or (Expressed in another way) to identify the presence of DNA specific to line 17053 in the target Dn a sample. ^ Provide a DNA molecule sufficient for use as a DNA probe that is suitable for the determination of 'detecting or identifying the presence or absence of DNA unique and unique to line 17053 DNA in a sample. The DNA molecule comprises at least 15 contiguous nucleotides of the ID N complex or a complement thereof, or SEq ID N〇: 2 of at least 15 contiguous nucleotides or a complement thereof. Nucleic acid amplification can be achieved by any of a variety of nucleic acid amplification methods known in the art, including thermal amplification methods. The heterologous DNA inserted into the rice line 17053 (including a representative seed sample of line 17053 registered with ATCC ρτΑ·9843) can be amplified by using primers derived from the sequences provided herein. The sequences of the line, followed by standard DNA sequencing of the amplicon or pure line DNA, are verified (corrected if necessary). The amplicons produced by these methods can be tested by a variety of techniques. One method is Genetic Bit Anaiysis (orov^^, Nucleic Acid Res. 22:4167-4175, 1994) « One of the designs - DNA nucleus, which is adjacent to the flanking genomic DNA 】 The inserted DNA sequence overlaps. The oligo-acid is immobilized in the wells of the microwell 147377.doc •33- 201040274 plate. After thermal amplification of the region of interest (a primer for the inserted sequence and an primer for the adjacent flanking sequence), the single-stranded amplicon (thermal amplification product) can The immobilized oligonucleotide hybridizes and serves as a template for a single base extension reaction using DNA polymerase and labeled ddNTPs specific for the next base expected. The results can be read based on fluorescence or ELISA. Detection of fluorescence or other signals is due to the presence of inserts/flanking sequences for successful amplification, hybridization, and single base extension. Another method is pyrosequencing technology as described by Winge (Innov. Pharma. Tech. 00: 1 8-24, 2000). In this method, a nucleus acid is designed which overlaps with the adjacent gene DNa and the inserted DNA joining sequence. Hybridizing the oligonucleotide to a single-stranded thermal amplification product (single-stranded amplicon) of the region of interest (a primer in the inserted sequence and a primer in the flanking sequence), and in the DNA Polymerase, ATP, sulfurylase, luciferase, apyrase, adenosine 5' phosphor-sulfate, and luciferin Cultivate in existence. Individual ddNTPs were added, which resulted in the detection of an optical signal. The light signal indicates the presence of the transgene insertion/flanking sequence due to successful amplification, hybridization, and single or multiple base extension.

The luminescence polarization described by Chen et al. (Genome Res. 9: 492-498, 1999) is a method that can be used to detect amplicons. Using this method, a nucleotide is designed which overlaps with the flank of the genomic DNA and the inserted DNA ligating sequence. Hybridization of the raised nucleotide with the single-stranded 147377.doc-34-201040274 amplification product of the region of interest (a primer in the inserted DNA and a primer in the flanking DNA sequence), and in the DNA Incubation in the presence of polymerase and fluorescently labeled ddNTPs. Single base extension results in the incorporation of ddNTPs. This incorporation can be measured as a change in polarization using a fluorometer. A change in polarization indicates the presence of a transgene insertion/flank sequence due to successful amplification, hybridization, and single base extension. TAQMAN® (PE Applied Biosystems, Foster City, CA) may also use the instructions provided by the manufacturer to detect and/or quantify the presence of DNA sequences. Briefly, a FRET oligonucleotide probe was designed which overlaps with the flanked DNA of the gene and the inserted DNA junction sequence. The FRET probe and the amplification primer (a primer in the inserted DNA sequence and one primer in the flanking gene sequence) are circulated in the presence of a thermostable polymerase and dNTP. Hybridization of the FRET probe results in cleavage and release of the fluorescent moiety on the FRET probe away from the quenching moiety. Fluorescent signals indicate the presence of flanking/transgenic gene insertion sequences due to successful amplification and hybridization. Molecular beacons for sequence detection have been described as described in Tyangi et al. (Nature Biotech. 14: 303-308, 1996). Briefly, a FRET oligonucleotide probe was designed which overlaps with the flanking DNA and the inserted DNA junction sequence. The unique structure of the FRET probe results in a secondary structure that maintains the fluorescent moiety in close proximity to the quenching moiety. The FRET probe and the amplification primer (a primer in the inserted DNA sequence and one primer in the flanking gene sequence) are circulated in the presence of a thermostable polymerase and dNTP. Upon successful amplification, hybridization of the FRET probe to the target sequence results in removal of the probe secondary structure and spatial separation of the fluorescent portion from the quenching moiety, resulting in a fluorescent signal. The fluorescent signal indicates the presence of a flanking/transgenic gene insertion sequence due to successful amplification and hybridization. Other methods are known in the art to be used in the practice of the present invention, such as: microfluidic technology (see, for example, U.S. Patent Publication No. 2006068398 and U.S. Patent No. 6,544,734), which provides separation and expansion. Method and apparatus for increasing DNA samples; optical dyes for detecting and measuring specific DNA molecules (see, for example, WO/05017181); nanotube devices comprising electronic sensors for detecting DNA molecules (see, for example, WO/06024023) And/or a nanobead of a specific DNA molecule that is subsequently measurable. DNA detection kits can be developed using the compositions disclosed herein and the Dna detection methods known in the art. These kits are suitable for breeding methods for identifying lines 1 7053 in a sample and for applying to rice plants containing appropriate strains of DNA. The kit may contain a DNA primer or probe or a fragment thereof that is similar or complementary to SEQ ID N: "6. The kit and detection method of the present invention are therefore particularly suitable for identifying rice lines 7053, selecting inclusion lines] 7〇 Plant variety or hybrid of 53, detected in the sample from the presence of the rice line 7〇53 and the presence and/or absence of the plant line, plant 4, or commodity for the rice line 17053 or the rice line containing 17〇53 The following examples are included to illustrate examples of certain preferred embodiments of the invention. Those skilled in the art will appreciate that the technology disclosed in the examples below represents that the inventors of the present invention have found that they have performed well in the practice of the present invention. The method, and thus may be considered as an example of a preferred mode of practice. However, it will be apparent to those skilled in the <RTIgt; Similar or similar results were obtained in the context of the spirit and scope of the present invention. Example 147377.doc • 36 - 201040274 Example 1: Transformation of rice and selection of strains This example describes how to form a gene transfer The colony line and how to select the line 17053. The gene-tolerant glyphosate-tolerant line 17053 is subjected to particle gun by using the DNA fragment of the gene shown in Figure 1 (the sequence of which is shown in SEQ ID NO: 5). The rice gene of the transgenic gene contains a representational cassette containing a promoter derived from Cauliflower Mosaic Virus (P-CaMV.e35S) containing a double fortifier. , operably linked to an intron molecule derived from the rice actin 1 gene (I-Os. Actl), operably linked to a chloroplast transit peptide (CTP2, iguanafen i/za/z_fl«a) DNA molecule of EPSPS), operably linked to a DNA molecule encoding glyphosate-resistant EPSPS (AGRtu.EPSPS: CP4), operably linked to a tumor-derived Agrobacterium tumefaciens (3 of Jgrokcie Wwm 胭 验 合成 synthase gene) Transcription termination region DNA molecule (T-AGRtu.nos). The rice variety M-202 explants were first transformed by particle gun method in one of four performance cassettes. Subsequently, the transformation was selected on a medium containing glyphosate. The cells and regenerate the viable cells. The process yielded 928 R0 plants, each of which was an independent, individual line. The 928 lines were screened by PCR and Southern analysis in the R0 phase to exclude multiple copies and/or molecular complex lines. In PCR analysis, the endpoint The TAQMAN® assay was initially used with the extracted DNA. Based on PCR screening, 565 R0 lines were selected. The 565 R0 lines were subsequently screened by Southern analysis. In the Southern analysis, DNA was extracted from rice tissue to Ncol and EcoRI. Or SspI digestion, and hybridization using radioactive DNA probes complementary to the CaMV 35S promoter 147377.doc-37-201040274 and EPSPS coding sequences using Southern blot hybridization analysis techniques. The data were used to determine the number of copies of the transgenic gene cassette inserted into the rice genome and to select 240 R0 lines with single insertion. The 240 selected lines were then promoted to growth chambers for phenotypic and fertilization analysis to identify off-type plants. Based on the growth box phenotype and fertilization rate screening, 1 70 lines were selected for promotion. The 1 70 R1 lines were then screened for glyphosate tolerance in a growth chamber. These plants were also analyzed for complete transgenic gene insertion by secondary Southern analysis. Using the data collected from these analyses, 19 lines were selected for promotion. Subsequently, 13 of the R2 plants in these lines were promoted to field trials. Plant field data on multiple features were collected and plants from each of the 13 lines were collected for one growing season, and five of them collected two growing seasons. In the first year of the field trial, glyphosate tolerance and agronomic equivalence of 13 lines were tested in a minimum of 3 positions in multiple replicate field trial designs. The results are presented in Table 1. Table 1: Field test results of the first season field trial plan Parameter line A 17053 BCDEFG Η IJ Κ L Effect Vegetative Tolerance Is it yes or yes is it yes or is it yield is yes or no Is it whether it is fertilization rate, phenotype, maturity? Is it whether it is yes or not? Is it agronomic yield? Is it yes or yes Yes Yes Yes Yes Yes Yes 147377.doc -38- 201040274 Fertilization Whether the rate, phenotype, maturity is whether it is yes or no, whether it is through field trials, whether it is whether it is whether or not it is subsequently used to select five leading lines to promote to the second year of the field. test. In these second year field trials, glyphosate tolerance and agronomic equivalence of five leading lines were tested in multiple replicate field trial designs at six locations. The results are presented in Table 2. Table 2: Field test results of the second season field trial plan Parameter line 17053 BCEG Efficacy Plant plantability is whether the yield is yes or not, the phenotype, the maturity is the agronomic yield Yes, is the fertilization rate, phenotype, maturity, whether it is through the field test, whether it is in the two seasons of efficacy test, in the growth period of 4-6 leaves with 3 pounds of acid equivalent / Plants are treated with acre (lb ae/ac) of glyphosate (which is twice the typical commercial ratio) or 4.5 lb ae/ac of glyphosate, which is three times the typical commercial ratio. The glyphosate plant tolerance was measured as a vegetative injury and the glyphosate reproductive tolerance was measured as yield and fertilization rate (%). Lines 1 7053 were selected using data from these trials. Line 17053 showed no vegetative damage at 3 lb ae/ac or 4.5 lb ae/ac and was measured in ton per acre (T/ac) of rice after treatment with glyphosate with no yield loss. 147377.doc -39- 201040274 Example 2: Isolation of rice chromosomal sequences adjacent to inserted DNA The genomic DNA for all PCR reactions was isolated using a rapid high pH/high salt lysis protocol. In this protocol, about 0.1 gram of lyophilized powdered leaf tissue is mixed with 600 ul (microliter) of lysis buffer (100 mM Tris, 1 Μ KC1, 10 mM EDTA, pH 9.5) by vortexing, followed by 65 Incubate at °C for 45-60 minutes. Next, the tube was vortexed again and 200 ul of precipitation buffer (5 Μ potassium acetate, pH 7.0) was added, vortexed again and centrifuged. An aliquot of 600 ul of DNA solution was transferred to a clean tube and 500 ul of ice-cold isopropanol was added to precipitate the DNA. After centrifugation, the DNA pellet was washed with 70% ethanol, air dried and the DNA resuspended in 250 ul of water. The extension of the 1 7053 line was obtained using a TAIL-PCR protocol as described in Liu et al. (Plant Journal 8: 457-463, 1995) to obtain DNA of the genomic DNA adjacent to the insertion point of the transgene. To identify flanking genomic DNA, two sets of two nested genome-walking primers were designed in the vicinity of the flanking junction. One set is designed to move from the 5' end of the integrated performance cassette and the other set is designed to move from the 3' end of the integrated performance cassette. The primer for 5' TAIL-PCR is the primer SQ1871 (SEQ ID NO: 12) in the first round and the primer SQ1869 (SEQ ID NO: 13) in the second round. 3. The primer for TAIL-PCR is the primer SQ1 875 (SEQ ID NO: 10) in the first round and the primer SQ1880 (SEQ ID NO: 14) in the second round.

In the identification of each flanking region, two nested primers were used in conjunction with a shorter random degenerate (AD) primer as described in Liu et al. (Plant Journal 8: 457-463, 1995) for continuous PCR reactions. . In the first round of PCR of the 5' and 3' flanking sequences, the corresponding PCR reactions were established as detailed in Table 3. At MJ 147377.doc -40- 201040274

The engine temperature cycler uses the cycle parameters shown in Table 4 and the preset ramp rate settings to perform these reactions. Table 3: The first round of TAIL-PCR reaction of rice line 17053

Step Reagent amount Remarks 1 18 MΩ water added to 50 ul final volume - 2 1〇χ Reaction buffer (containing MgCl2) 5.0 ul lx buffer final concentration, 1.5 mM MgCl2 final concentration 3 10 mM dATP, dCTP, dGTP and dTTP Solution 1.0 ul 200 μΜ each dNTP final concentration 4 primer SQ1871 (5' side) or primer SQ1875 (3' side) (resuspended in 1 χΤΕ buffer or 18 mega ohms to ΙΟμΜ) 1.0 ul 0.2 μΜ Final concentration 5 Random degenerate primers ADI, AD2 or AD3 (resuspended in 1 χΤΕ buffer or 18 mega ohms water to 1 〇〇μΜ concentration) 1.5 ul 3.0 μΜ final concentration It is recommended to use all three random degenerate primers for maximum Success rate 6 REDTaq DNA polymerase (1 unit / ul) 2 · 5 ul (recommended to convert the pipette before the next step) 2.5 units / reaction 7 extracted DNA (template): target DNA positive control DNA non-gene transgenic DNA 50 -200 ng of genomic DNA 1) · It is recommended to use high-quality DNA as a template to increase the success rate. 2) It is recommended to use the positive control DNA from the gene-transferred line whose flank sequence has been successfully isolated using this method. Doc -41 - 201040274 Table 4: 1st round of TAIL-PCR thermal cycling conditions Cycle number setting 1 94〇C 2 minutes 5 94〇C 30 seconds 62〇C 1 minute 72〇C 2 minutes 30 seconds 1 94〇C 30 seconds 25〇C 3 minutes, 3 minutes to 72〇c (〇2〇c/sec) 72°C 2 points 3〇ίΨ 15 94〇C 10 seconds 68〇C 1 minute 72〇C 2 minutes 30 seconds 94〇 C 10 seconds 68〇C 1 minute 72〇C 2 minutes 30 seconds 94〇C 10 seconds 44〇C 1 minute 72〇C 2 minutes 30 seconds 1 72〇C 5 minutes The first round of TAIL-PCR aliquot For the first round of TAIL-PCR as detailed in Table 5. These reactions were performed in the MJ Engine temperature cycler using the cycle parameters shown in Table 6 and the preset ramp rate settings. Table 5: Stage 2 TAIL-PCR reaction step Reagent amount Remarks 1 18 MΩ water added to 50 ill Final volume - 2 l〇x reaction buffer (containing MgCl2) 5.0 ul 1χ buffer final concentration, 1.5 mMMgCl2 final concentration 3 10 mM dATP, dCTP, dGTP and dTTP solutions 1.0 ul 200 μΜ each dNTP final concentration 147377.doc -42- 201040274 4 primer SQ18690 side) or primer SQ1880 (3' side) (resuspended in 1 χΤΕ buffer) Or a concentration of 18 megohms to 10 μΜ) 1.0 ul 0.2 μΜ final concentration 5 random degenerate primer (resuspended in 1 x TE buffer or 18 meg water to 100 μΜ) 1.0 ul 2.0 μΜ final concentration 6 REDTaq DNA Polymerase (1 unit / ul) 2.5 ul 2.5 units / reaction (recommended to switch the pipette before the next step) 7 1:100 dilution of the first round of TAIL-PCR reactant 5.0 ul Table 6: The second round of TAIL-PCR heat The cycle condition cycle number is set to 1 94. . 2 minutes 12 94〇C 10 seconds 64〇C 1 minute 72〇C 2 minutes 30 seconds 94〇C 10 seconds 64〇C 1 minute 72〇C 2 minutes 30 seconds 94〇C 10 seconds 44〇C 1 minute 72〇C 2 minutes 30 seconds 10 94 〇 C 15 seconds 44 〇 C 1 minute 72 ° C 2 minutes 30 seconds 1 72 〇 C 5 minutes

To observe the TAIL-PCR amplicon, an aliquot of the second round of reaction was run on a 2.0% agarose gel and stained with ethidium bromide. In addition, an aliquot of the second round of TAIL-PCR was sent to the Monsanto genomic sequencing centering sequence. Sequence analysis was performed using Monsanto's proprietary Flanking Sequence Application sequence analysis software. The PCR primers were designed to confirm the base observed in the sequencing results of the second round of the TAIL-PCR reaction. 147377.doc -43- 201040274 Factor-directed sequence. The 5' genomic flanking sequence for line 17053 was confirmed by PCR reaction using primer pair SQ3626 (SEQ ID NO: 15) and SQ1869 (SEQ ID NO: 13). The 3' genobody flanking sequence for line 17053 was confirmed by PCR reaction using primer pair SQ3 623 (SEQ ID NO: 11) and SQ1875 (SEQ ID NO: 10). Example 3: Line-specific endpoint TAQMAN® assay This example describes the TAQMAN® thermal amplification method for the identification of the specific end point of the strain developed in the sample 1 7 5 5 3 . Examples of conditions suitable for this method are described in Tables 7 and 8. Suitable DNA molecules for this method are, for example, the primers SQ4194 (SEQ ID NO: 7) and SQ4191 (SEQ ID NO: 8) and the 6FAMtm-labeled oligonucleotide probe PB1494 (SEQ ID NO: 9). Other probes and primers can be designed based on the sequences of the transgene insertion sequences and/or flanking sequences provided herein. SQ4194 (SEQ ID NO: 7) and SQ4191 (SEQ ID NO: 8), when used in conjunction with PB1494 (SEQ ID NO: 9) for such reaction methods, produce a DNA amplicon of the identified line 17053 DNA. Controls for this analysis included a positive control from rice containing line 17053 DNA, a negative control from non-genegenic rice, and a negative control without template DNA. These assays were optimized for use with the Applied Biosystems GeneAmp PCR System 9700 or Stratagene Robocycler, MJ Engine, Perkin-Elmer 9700 or Eppendorf Mastercycler Gradient. Other methods and devices are known to those skilled in the art that will be suitable for generating the amplicon of line 17053 DNA in a biological sample. When analyzing samples, the cycle parameters shown in Tables 7 and 8 can be used. At 147377.doc -44- 201040274

When performing thermal amplification in an Eppendorf Mastercycler Gradient or MJ Engine, the temperature cycler should be operated in calculation mode. When performing thermal amplification in perkin_Elmer 9700, the temperature cycler should be set to maximum uniformity. Table 7: Rice Line 17053 Specific Endpoint TAQMAN® Step Reagent Volume Remarks 1 18 MΩ Water Adjusted to 10 μΐ Final Volume 2 2x Universal Standard Mix (dNTP, Enzyme, Buffer) 5.0 μΐ 1χ Final Concentration 3 Primer-1 and Primer-2 Mixture (resuspended in 18 MΩ water to a concentration of 20 uM) Example: In a microcentrifuge tube, the following should be added to achieve a concentration of 500 μΐ at a final concentration of 20 μM: 100 μΜ 100 μΐ primer SQ4194 (SEQ ID NO: 7); 100 μΜ concentration of 100 μΐ primer SQ4191 (SEQ ID NO: 8); 300 μ118 Meg water 0.5 μΐ 1.0 μΜ final concentration 4 strain 6-FAMTM MGB probe PB1494 (SEQ ID NO: 9) (Resuspend in 18 MΩ water to 10 μΜ concentration) 0.2 μΐ 0.2 μΜ Final concentration 5 Extracted DNA (template>: 1. Leaf sample to be analyzed 2. Negative control (non-gene transfer) DNA) 3. Negative water control (no template) 4. Positive control 17053 DNA 3.0 μΐ Table 8: End point TAQMAN® Thermal cycler Condition cycle number setting 1 50°C 2 minutes 1 95〇C 1〇10 10〇C 15 Seconds 64t 1 minute 147377.doc • 45- 201040274 -l °c / cycle 30 95 ° C 15 seconds 54V 1 minute 1 10 ° C Permanent Example 4: Identification of lineage 1753 in terms of breeding activity This example describes how a child with line 1 7053 can be used in a child with any breeding activity Internal recognition line 1 7053. An amplicon of the identification line 17053 was generated using a DNA line primer pair. The amplicon of the identification line 17053 comprises at least one SEQ ID NO: 1 or SEQ ID NO: 2 ([A] and [ B], the ligation sequence provided in Figure 1 respectively. SEQ ID NO: 1 ([A] of Figure 1) corresponds to the flanking sequence and the 5' end of the transgene insertion sequence (SEQ ID NO: 3) The position of [C] is 565 to 5 84, see the nucleotide sequence of the junction sequence of Figure 1). SEQ ID NO: 2 ([B], see Figure 1) corresponds to the flanking sequence and the transgene insertion sequence 3 The nucleotide sequence of the junction sequence of the 'end (SEQ ID NO: 4 [D], 626 to 645, see Figure 1). The line primer pair that will produce the identified amplicon of line 17053 includes the use of flanking sequences (SEQ ID NO: 3 and 4) and the primer pair designed for the inserted gene transfer DNA sequence (SEQ ID NO: 5). To obtain an identified amplicon comprising at least 11 nucleotides of SEQ ID NO: 1, a pre-priming molecule based on bases 1 to 564 of SEQ ID NO. 3 is designed and based on the inserted expression cassette DNA The inverted primer molecules of positions 1 to 3142 of SEQ ID NO: 5, wherein the primer molecules have contiguous nucleotides of sufficient length to specifically hybridize to SEQ ID NO: 3 and SEQ ID NO: 5. To obtain an identified amplicon containing at least 11 nucleotides of SEQ ID NO: 2, a pre-priming based on position 1 to 3142 of the inserted expression cassette SEQ ID NO: 5 is designed to be 147377.doc-46 - 201040274 and an inverted primer molecule based on bases 1 to 645 of the 3' flanking sequence SEQ ID NO: 4 wherein the primer molecules have contiguous nucleotides of sufficient length to SEQ ID NO: 4 and SEQ ID NO: 5 specific hybridization. For practical purposes, primers should be designed to produce amplicons with a limited range of sizes (e.g., between i 〇〇 and 丨〇〇〇 bases). Amplicon of smaller size (shorter length of polynucleotide) is generally more reliable and can be generated in PCR reactions that allow for shorter cycle times, and is easily separated and observed on an agarose gel or suitable for the endpoint. TAQMAN® class certification. Smaller amplicons can be generated and detected by DNA amplicon detection methods known in the art. In addition, the amplicon produced using the primer pair can be selected for propagation into the vector, propagated, isolated and sequenced, or can be directly sequenced using methods well defined in the art. Any amplicon derived from the combination of SEQ ID NO: 3 and SEQ ID NO: 5 or the combination of SEQ ID NO: 4 and SEQ ID NO: 5 and suitable for use in a DNA amplification method to produce the identification line 17053 or a progeny thereof The introduction of the pair is an aspect of the invention. Any single, isolated DNA polynucleus comprising at least 11 contiguous ribonic acids of SEQ ID NO: 3 or a complement thereof and suitable for use in a DNA amplification method to produce an amplicon identifying a plant or a progeny thereof comprising line 17053 The glucoside primer molecule is one aspect of the invention. Any single, isolated, at least 11 contiguous nucleotides of SEQ ID NO: 4 or the complement thereof and suitable for use in a DNA amplification method to produce a DNA polynucleus that identifies an amplicon comprising a plant or a progeny of line 17053 The glucoside primer molecule is one aspect of the invention. Any single, isolated DNA comprising at least 11 contiguous nucleotides of SEQ ID NO: 5 or a complement thereof and suitable for use in a DNA amplification method to produce an amplicon identifying a plant or a progeny thereof comprising line ι7〇53 The polynucleotide primer molecule is one of the aspects of the invention 147377.doc -47· 201040274. Examples of amplification conditions used in this analysis are illustrated in Tables 7 and 8. However, such methods are either DNA primers homologous or complementary to the DNA sequence of the transgene insertion of SEQ ID NO: 3 or SEQ ID NO: 4 or line 17053 (SEQ ID NO: 5) to generate amplification of the identification line 17053. Any improvement in the usage of the child is within the scope of the invention. The amplicon is identified to comprise a DNA molecule that is homologous or complementary to at least one of the transgene/genomic junction DNA (SEQ ID NO: 1 or SEQ ID NO: 2) or a substantial portion thereof. Analysis of line 17053 in the sample can include a positive control from line 17053, a negative control from a non-lineage 17053 (e.g., but not limited to, M-202) rice plants, and/or a negative control containing no rice genomic DNA. The pair of primers that amplify the endogenous rice DNA molecule can serve as an internal control for DNA amplification conditions. Any sequence fragment selected from the sequence set forth in SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5 can be used as a DNA amplification primer for generation by the methods shown in Tables 7 and 8. The amplicon, and when the line 1 7053 was used as a template to identify the amplification reaction, the amplicon can identify line 17053. Improvements using the DNA primer sequences and the methods of Tables 7 and 8 are within the scope of the present invention. An amplicon for identifying line 17053 produced by at least one DNA primer sequence derived from S E Q ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5 is one aspect of the invention. The DN A detection kit can therefore be designed to contain at least one DNA primer having a sufficient length contiguous nucleotide derived from SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5 and used for DNA amplification An increase in the method yields an identification 147377.doc -48- 201040274 An amplicon comprising a plant of line 1 7053 or a progeny thereof, and which is an aspect of the invention. A rice plant part or seed or commodity which produces an amplicon identifying line 17053 when tested in a DNA amplification method is one aspect of the invention. The assay for line 17053 amplicons can be performed as shown in Table 8 by using Applied Biosystems GeneAmp® PCR System 9700 or

Stratagene RoboCycler® or MJ Engine or Perkin_Elmer 97(R) or Eppendorf Mastercycler® Gradient Temperature Cycler or any other amplification system that can be used to generate the amplicons of the identified line 17053 is included in the scope of the above-disclosed patent application. A representative seed sample of the rice line 17〇53 has been deposited with the American Type Culture Collection (ATCC) under the Budapest Treaty (University Avenue, Manassas, VA, USA), zip code 20110). The registration date is 2_years and 2 days. The ATCC registration number for this deposit is PTA_9843. The deposit will remain in the deposit (four) year period, or the five-year period after the last request, or the patent expiration date (take a long period of time) and can be replaced if needed during that time. The principles of the present invention have been illustrated and described, but it will be apparent to those skilled in the art that the present invention may be practiced without departing from the scope of the principles. The spirit of the stomach (four) and the spirit of the circle [simplified description of the schema] Figure 1 shows the diagram of the rice line 17053: [Α position (as provided in SEQ ID NO: 1), the relative cause of the μ mouth &amp; The 5th part of the cholestyramine is the junction between the ungenerated body and the inserted raft, and [B] corresponds to the relative position of the 3' junctional region 147377.doc •49· 201040274 (eg SEQ ID NO: 2) Provided), which is the junction between the rice genome and the 3' portion of the inserted gene-transferred DNA; [C] corresponds to the 5' flanking region and the relative 5' end of one of the inserted gene-transferred DNA Position (as provided by SEQ ID NO: 3), which comprises the 5' rice genome sequence of the integrated expression cassette of the flanking line 17053 and the 5' end region of the transgenic DNA; [D] corresponds to 3 The relative position of the flanking region and the 3' end of one of the inserted gene-transforming DNA (as provided by SEQ ID NO: 4), which includes the integrated expression of the flanking line 17053, the 3'-m gene sequence of the cassette And the region of the 3' end of the transgenic DNA; [E] indicates the transgene expression of the inserted line 17053 gene (eg SEQ ID) NO.·5 provides); and [F] represents the contiguous sequence of the flanking sequence and the transgene expression cassette (as provided by SEQ ID NO: 6), which contains the SEQ ID from left to right as shown in the figure. NO: 3, SEQ ID NO: 5 and SEQ ID NO: 4, wherein SEQ ID NO: 1 and SEQ ID NO: 2 are incorporated as described above due to the junction sequence representing line 17053. 147377.doc -50- 201040274

序列 Sequence Listing &lt;110&gt; American Monsanto Technology Co., Ltd. &lt;120 Gene Transgenic Rice Line 17053 and Method of Use &lt;130&gt; 38-21(56582)0001 &lt;140&gt; 099109684 &lt;141&gt; 2010/03 /30 &lt;150&gt; 61/164,899 &lt;151&gt; 2009-03-30 &lt;160&gt; 15 &lt;170&gt; Patentln version 3.5 &lt;210&gt; 1 &lt;211&gt; 20 &lt;212&gt;&lt;213&gt; DNA artificial Sequence &lt;220&gt;&lt;223&gt; chemically synthesized oligonucleotide; chimeric DNA molecule of rice gene DNA and transgenic DNA &lt;400&gt; 1 aaaggataca accaagcttg Z0 &lt;210&gt; 2 &lt;211&gt; 20 &lt;;212&gt;DNA.&lt;213&gt;Artificialsequence&lt;220&gt;&lt;223&gt; Chemically synthesized oligonucleotide; chimeric DNA molecule of rice gene DNA and transgenic DNA&lt;400&gt; 2 tagtctatgt taagcttggt 20 &lt;;210&gt; 3 &lt;211&gt; 584 &lt;212&gt; DNA artificial sequence &lt;220&gt;&lt;223&gt; rice genomic DNA and chimeric DNA molecule of transgenic DNA&lt;400&gt; 3 ttgtagaagt taagcaaaag tcaatggcat attgtagaac dtgataaagt cagtgjgcaaa tcgtagacgt Gtgacaaaat cagtggtata taatggattc tctctattat tt tagaccta tgattgtctt gcagtaagat agtgtagaag tcttggtcat tgcgagctgc tcgctttttg tcattgatgt agcattacat tgtttagtca agaagtactg ctgctcgctc gagtcattca 60 120 180 240 147377.doc 201040274 attcgttaga cgttgcttct gattgaatca tgcatgaagt acctatctgt aaccaatgtt 300 attctagatg gtagacaata aatattggat caccgtttgt ttacttgtta tacttgacaa 360 atagacacag aacgtgacaa acatcggata ccatgtccag tatgcatgat aatttaaaaa 420 aagaacaatc tagctcatca ccccggtcaa aatatatgct ttaatttctt acaagattaa 480 tcctatcttg Tgaggaggca ggtggatcga tccatcggag ttaattaa&lt;ia agactaagga 540 agaccataat agagaatcca aataaaagga tacaaccaag cttg 584 &lt;210&gt; 4 &lt;211&gt; 645 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Rice genomic DNA and transformation the chimeric DNA molecule of genomic DNA &lt; 400 &gt;.. 4 caaaagtaac tggcggcgtg gccatcgcta tagaaagaaa gaaggcacca gggtgtcgtc 60 · · · · ggcaacagag gtggccagct tgtcatcgcg gtcagagtcg gagcttcacg tcattgccgc 120 gacggtggtt gggctctcag tctccatgtt actgcctggg gaagacgggt tcgacgacat 180 ggatgaggag gagagcagcg gcggcgtgtg ctaacgagga ggaggaaaga gagagatgat 240 gaagggttag aggatgggaa agaggaaggg ggagaggagg ggaaagagag gggtgttgac 300 gtgggttcca caatgttttt ttttactata taagtgccac gtcaacacta tataggtgtt 360 acatcagtcg aaactgaggg ggtgtttgga tgggactgaa actctttagt ccctgtcaca 420 tcggatgttt gaacaccaat tagaagtatt aaacgtagac taatgacaaa acccattcta 480 taaccctaga ctgattcaca ctacgaatct atgtgagcct aatttatcaa tgattagcct 540 atgtgatgct acagtaaata ttctctcata tggattaatt agtcttaaaa aatttgtctc 600 gcgaattagc tctcatttat gtaagtagtc tatgttaagc ttggt 645 &lt; 210 &gt; 5 &lt; 211 &gt; 3142 &lt; 212 &gt; DNA &lt; 213 &gt; artificial sequence &lt; 220 &gt; accaagcttg atatccctag ggcggccgcg ttaacaagct tctgcaggtc cgattgagac 60 ttttcaacaa agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca 120 ctttattgtg aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa 180 aggaaaggcc atcgttgaag Atgcctctgc cgacagtggt cccaaagatg gacccccacc 240 &lt;223&gt; chemically synthesized oligonucleotide; chimeric DNA molecule of rice gene DNA and transgenic DNA &lt;400&gt; 5 147377.doc 2-201040 274 cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc siagtggattg atgtgatggt ccgattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt ccattgccca gctatctgtc acttt'attgt gaagatagtg will gtg will cttcta. Caaatgccat cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg. · .. .. "... tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaiaccac •, .. ;.. .., gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc ...- '' ccactatcct tcigcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac acgctgacaa gctgactcta gcagatcctc tagaaccatc ttccacacac tcaagccaca ctattggaga acacacaggg acaacacacc ataagatcca agggaggcct ccgccgccgc cggtaaccac cccgcccctc tcctctttct ttctccgttt ttttttccgt ctcggtctcg atctttggcc ttggtagttt giggtgggcga gaggcggctt cgtgcgcgcc cagatcggtg cgcgggaggg gcgggatctc Gcggctgggg ctctcgccgg cgtggatccg gcccggatct 〇cgcggggaat ggggctctcg gatgtagatc tgcgatccgc cgttgttggg ggagatgatg gggggtttaa aatttccgcc gtgctaaaca agatcaggaa gaggggaaaa gggcactatgi . · gtttatattt t tatatattt ctgctgcttc gtcaggctta gatgtgctag atctttcttt cttctttttg tggigtagaat ttgaatccct cagcattgtt catcggtagt ttttcttttc •: ·· · '· atgatttgtg acaaatgcag cctcgtgcgg agcttttttg taggtagaag tgatcaa ^ ca; tggcgcaagt tagcagaatc tgcaatggtg tgcagaaccc atctcttatc tccaatctct - * · · cgaaatccag tcaacgcaaa tctcccttat cggtttctct gaagacgcag cagcatccac gagcttatcc gatttcgtcg tcgtggggat tgaagaagag tgggatgacg.. ttaattggct ctgagcttcg tcctcttaag gtcatgtctt ctgtttccac ggcgtgcatg ctacacggtg caagcagccg gccggcaacc gctcgcaaat cttccggcct ttcgggaacg gtcaggattc cgggcgataa gtccatatcc caccggtcgt tcatgttcgg cggtcttgcc agcggtgaga ^ cgcgcatcac gggcctgctt gaaggtgagg acgtgatcaa taccgggaag gccatgcagg ctatgggagc gcgtatccgc aaggaaggtg acacatggat cattgacggc gttgggaatg. gcggtctgct cgcccctgag gcccctctcg acttcggcaa tgcggcgacg ggctgcaggc tcactatggg actggtcggg gtgtacgact tcgatagcac gttcatcgga gacgcetcgc tcacaaagcg cccaatgggc cgcgttctga acccgttgcg cgagatgggc Gtacaggtca aatccgagga tggtgaccgt ttgcccgtta cgctgcgcgg gccga agacg cctaccccga ttacctaccg cgtgccaatg gcatcegccc aggtcaagtc agccgtgctc ctcgccggac tgaacactcc gggcatcacc acggtgatcg agcccatcat gaccagggat cataccgaaa agatgcttca ggggtttggc gccaacctga cggtcgagac ggacgctgac ggcgtcagga 147377.doc 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 H4Q 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 201040274 ccatccgcct tgagggcagg ggtaaactga ctggccaagt catcgatgtt ccgggagiacc cgtcgtccac ggccttcccg ttggttgcgg cgctgctcgt gccggggagt gacgtgacca tcctgaacgt cctcatgaac ccgaccagga ccggcctgat cctcacgctt caggagatgg •.. · · · ·. gagccgacat cgaggtgatc aacccgcgcc tggcaggcgg tgaagacgtt gcggatctgc.. ..... gcgtgcgctc ctctaccctg aagggcgtga cggtcccgga agatcgcgcg ccgtccatga tagacgagta tcctattctg gccgtcgccg ctgcgttcgc cgaaggggcc acggtcatga acggtcttga ggaactccgc gtgaaggaat cggatcgcct gtcggcggtg gccaatggcc tgaagctcaa cggtgttgac tgcgacgagg gtgagacctc actcgtggtc cgtggccggc - · ctgatggcaa gggcctcggc aacgccagtg gagcggccgt cgcca cgcac ctcgatcatc gcatcgcgat gtccttcttg gtgatgggtc tcgtctcaga gaacccggtg accgtcgatg • · acgccacgat gatagcgacg agcttcccag agttcatgga tctgatggcg ggcctcgggg ccaagatcga actgtctgac acgaaggccg cttgaattcc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcggggat ggggaattcg gtaccaagct ta &lt;210&gt; 6 &lt;211&gt; 4351 &lt;2X2&gt; DNA &lt;213&gt; artificial sequence. nucleotides for chemical synthesis; host DNA DNA of rice gene and transfer gene DNA &lt;400&gt; 6 ttgtagaagt taagcaaaag tcaatggcat attgtagaac atgataaagt cagtggcaaa tcgtagacgt gtgacaaaat cagtggtata taatggattc tctctattat tttagaccta '. tgattgtctt gcagtaagat agtgtagaag tcttggtcat tgcgagctgc tcgctttttg tcattgatgt agcattacat tgtttagtca agaagtactg ctgctcgctc gagtcattca attcgttaga cgttgcttct gattgaatca tgcatgaagt acctatctj gt aaccaatgtt attctagatg gtagacaata aatattggat caccgtttgt ttiacttgtta tacttgacaa atagacacag aacgtgacaa acatcggata.ccatgtccag tatgcatgat.aatttaaaaa aagaacaatc tagctcatca ccccggtcaa aatatatgct ttaatttctt acaagattaa tcctatcttg tgaggaggca ggtggatcga tccatcggag ttaattaaaa agactaagga 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3142 60 120 180 240 300 360 420 480 540 147377.doc -4- 201040274 agaccataat agagaatcca aataaaagga tacaaccaag cttgatatcc ctagggcggc 1 . . . ... . cgcgttaaca agcttctgca ggtccgattg agacttttca acaaagggta atatccggaa ' . . . . . . ...:: acctcctcgg attccattgc ccagctatct gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt gaagatgcct · · · .. · ... ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaaigaag: -...... · · · ·, · acgttccaac cacgtcttca aagcaagtgg attgatgtga tggtccgatt gagacttttc · · aacaaagggt aatatccgga aacctcctcg Gattccattg cccagctatc tgtcacttta ttgtgaagat agt ggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa round .. aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg &quot; · atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct - 'ctatataagg aagttcattt catttggaga ggacacgctg acaagctgac tctagcagat .... · Ο 〇cctctagaac c...? tcttccac acactcaagc caeactattg gagaacacac agggacaaca •. · ·. · caccataaga tccaagggag gcctccgccg ccgccggtaa ccaccccgcc cctctcctct ttctttctcc gttttttttt ccgtctcggt ctcgatcttt ggccttggta gtttgggtgg gcgagaggcg gcttcgtgcg cgcccagatc ggtgc ^ cggg aggggcggga tctcgcggct ggggctctcg ccggcgtgga tccggcccgg atctcgcggg gaatggggct ctcggatgta gatctgcgat ccgccgttgt tgggggagat gatggggggt ttaaaatttc cgccgtgcta aacaagatca ggaagagggg aaaagggcac tatggtttat atttttatat atttctgctg. · ; . . -- ,..- cttcgtcagg cttagatgtg ctagatcttt ctttcttctt tttgtgggta gaatttgaat ccctcagcat tgttcatcgg tagtttttct tttcatgatt tgtgacaaat geagcctcgt gcggag cttt tttgtaggta gaagtgatca accatggcgc aagttagcag aatctgcaat ggtgtgcaga acccatctct tatctccaat ctctcgaaat ccagtcaacg caaatctccc ttatcggttt ctctgaagac gcagcagcat ccacgagctt atccgatttc gtcgtcgtgg ggattgaaga agagtgggat gacgttaatt ggctctgagc ttcgtcctct taaggtcatg tcttctgttt ccacggcgtg catgctacac ggtgcaagca gccggccggc daccgctcgc. aaatcttccg gcctttcggg aacggtcagg attccgggcg ataagtccat atcccaccgg tcgttcatgt tcggcggtct tgccagcggt gagacgcgca tcacgggcct gcttgaai ^ gt gaggacgtga tcaataccgg gaaggccatg caggctatgg gagcgcgtat ccgcaaggaa • - - · ggtgacacat ggatcattga cggcgttggg aatggcggtc tgctcgcccc tgaggcccct ctcgacttcg gcaatgcggc gacgggctgc aggctcacta tgggactggt cggggtgtac 147377.doc 201040274 gacttcgata gcacgttcat cggagacgcc tcgctcacaa agcgcccaat gggccgcgtt 2460 ctgaacccgt tgcgcgagat gggcgtacag gtcaaatccg aggatggtga ccgtttgccc ..2520 • · · -... · gttacgctgc gcgggccgaa gacgcctacc ccgattacct accgcgtgcc Aatggcatcc 2580 gcccaggtca agtcagccgt gctcctcgcc ggactgaaca ctccgggcat caccacggtg 2640 - . - . atcgagccca tcatgaccag ggatcatacc gaaaagatgc ttcaggggtt tggcgccaac 2700

I ctgacggtcg agacggacgc tgacggcgtc aggaccatcc gccttgaggg caggggtaaa 2760 ctgactggcc aagtcatcga tgttccggga gacccgtcgt ccacggcctt cccgttggtt 2820 ...., gcggcgctgc tcgtgccggg gagtgacgtg accatcctga acgtccteat gaacccgacc 2880 aggaccggcc tgatcctcac gcttcaggag atgggagccg acatcgaggt gatcaacccg 2940 cgcctggcag gcggtgaaga cgttgcggat etgcgcgtgc gctcctctac cctgaagggc 3000 gtgacggtcc cggaagatcg cgcgccgtcc atgatagacg agtatcctat tctggccgtc 3060 gccgctgcgt tcgccgaagg ggccacggtc atgaacggtc ttgaggaact ccgcgtgaag 3120 gaatcggatc gcctgtcggc ggtggccaat ggcctgaagc tcaacggtgt tgactgcgac 31B0 gagggtgaga cctcactcgt ggtccgtggc cggcctgatg gcaagggcct cggcaacgcc 3240 •. · agtggagcgg ccgtcgccac gcacctcgat catcgcatcg cgatgtcctt cttggtgatg 3300 ggtctcgtct cagagaaccc ggtgaccgtc gatgacgcca cgatgatagc gacgagcttc 3360 ccagagttca tggatctgat ggcgggcctc ggggccaaga tcgaactgtc tgacacgaag 3420 gccgcttgaa ttcccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct 3480 Gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcat gtaata 3540 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa 3600. ·. ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg 3660 cgcgcggtgt catctatgtt actagatcgg ggatggggaa ttcggtacca agcttaacat 3720 • ... agactactta cataaatgag agctaattcg cgagacaaat tttttaagac taattaatcc 3780 atatgagaga atatttactg tagcatcaca taggctaatc attgataaat taggctcaca 3840 tagattcgta gtgtgaatca gtctagggtt atagaatggg ttttgtcatt agtctacgtt 3900 taatacttct aattggtgtt caaacatccg atgtgacagg gactaaagag tttcagtccc 3960 atccaaacac cccctcagtt tcgactgatg taacacctat atagtgttga cgtggcactt 4020 atatagtaaa aaaaaacatt gtggaaccca cgtcaacacc cctctctttc ccctcctctc 4050 ccccttcctc tttcccatcc tctaaccctt catcatctct ctctttcctc ctcctcgtta 4140 gcacacgccg ccgctgctct cctcctcatc catgtcgtcg aacccgtctt ccccaggcag 4200 taacatggag actgagagcc caaccaccgt cgcggcaatg acgtgaagct ccgactctga 4260 ccgcgatgac aagctggcca cctctigttgc cgacgacacc ctggtgcctt ctttctttct 4320 147377.doc -6- 201040274 atagcgatgg ccacgccgcc agt Tactttt g &lt;210&gt; 7 &lt;211&gt; 28 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Chemically synthesized oligonucleotide PCR primer &lt;400&gt;

Taaggaagac cataatagag aatccaaa &lt;210&gt; 8 &lt;211&gt; 22 &lt;212&gt;DNA;&lt;213&gt;. Artificial sequence &lt;220&gt;

&lt;223&gt; Chemically synthesized oligonucleotide PCR primer &lt;400&gt; 8 gacctgcaga agcttgttaa eg &lt;210&gt; 9 &lt;211&gt; 17 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Chemically synthesized oligonucleotide PCR primer &lt;400&gt; 9 aggatacaac caagctt

&lt;210&gt; 10 &lt;211&gt; 25 &lt;212&gt; DNA i&lt;213&gt; 'Artificial sequence&lt;220&gt;&lt;223&gt; Chemically synthesized oligonucleotide PCR primer&lt;400&gt; 10 egettgaatt cccgatcgtt caaac &lt;210&gt 11 &lt;211&gt; 24 &lt;212&gt; DNA &lt;Π3&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Chemically synthesized oligonucleotide PCR primer &lt;400&gt; 11 ggctctcagt ctccatgtta ctgc 147377.doc 201040274 &lt;210&gt Ϊ2 &lt;211&gt; 24 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Chemical synthesis of nucleotide primers &lt;400&gt; 12 gaagacgtgg ttggaacgtc ttct &lt;210&gt; 13 &lt; 211 &gt; 24 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Chemically synthesized oligonucleotide PCR primer &lt;400&gt; 13 cggaccatca catcaatcca cttg &lt;210&gt; 14 &lt;211&gt; 24 &lt;;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Chemically synthesized oligonucleotide PCR primer &lt;400&gt; 14 gattgaatcc tgttgccggt cttg &lt;210&gt; 15 &lt;211&gt; 24 &lt;212&gt; DNA &lt;213&gt;Artificial sequence &lt;220&gt;&lt;223&gt; Nucleotide PCR primers &lt; 400 &gt; 15 cagtggcaaa tcgtagacgt gtga 147377.doc

Claims (1)

201040274 VII. Patent Application Range: 1. A DNA molecule comprising: a. having a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. a polynucleotide molecule; b. a polynucleotide molecule having the sequence of at least 15 contiguous nucleotides of nucleotide positions 565-584 of SEQ ID NO: 3, c· having the nucleus of SEQ ID NO: A polynucleotide molecule having a sequence of at least 11 contiguous nucleotides at positions 626-645; or d_ complementary to the sequence of (a), (b) or (c). 2. The DNA molecule of claim 1, which comprises at least 90% of SEQ ID NO: 6. 3. The DNA molecule of claim 1 wherein the DNA molecule is derived from an event 17053. 4. The DNA molecule of claim 1, wherein the DNA molecule is contained in a rice plant, plant cell, seed, progeny plant, plant part or commodity comprising line 17053, and a representative seed sample comprising line 17053 is deposited at the ATCC Registration number PTA-9843. 5. The DNA molecule of claim 1, wherein the DNA molecule is an amplicon produced by a template molecule of line 17〇53. 6. A polynucleotide probe for the identification of line 17053, wherein the probe is of sufficient length to bind at least 11 contiguous nucleus of a sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2. The nucleotide sequence and its complement. a DNA molecule consisting of a first DNA molecule and a second DNA molecule different from the first DNA 147377.doc 201040274 molecule, wherein the DNA molecule has a sufficient length of contiguous nucleotides of SEQ ID NO: 6. The nucleotide sequence or its complement, when used in conjunction with the template for rice line 17053 for amplification reactions, serves as a DNA primer to generate an amplicon of the rice line 17053 in the identified sample. 8. The DNA molecule pair of claim 7, wherein the amplicon comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2, and a complement thereof. 9. A method of detecting the presence of a DNA molecule of rice line 17053 comprising: a. contacting a DNA sample with a pair of DNA molecules as claimed in claim 7; b. performing an amplification reaction sufficient to produce an amplicon, the amplicon a polynucleotide molecule comprising at least 11 contiguous nucleotides selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2, and complementary sequences thereof; and c. detecting the DNA amplicon, Wherein the detection of the DNA amplicon identifies the presence of the DNA molecule of the rice line 1705 3 in the DNA sample. 10. A method of detecting the presence of a DNA molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 3 or SEQ ID NO: 4, comprising: a. contacting the sample with an inclusion selected from the group consisting of ID NO: 1 and a DNA probe of the nucleotide sequence of the group consisting of SEQ ID NO: 2 and a complementary sequence thereof, wherein the DNA probe is under stringent hybridization conditions and comprises a SEQ ID NO: selected from 147377.doc 201040274 SEQ ID NO: 3 or a DNA molecule of the nucleotide sequence of the group consisting of SEQ ID NO. 4 hybridizes and does not comprise a core selected from the group consisting of SEQ ID NO: 3 or SEQ ID NO: 4 under such stringent hybridization conditions DNA molecule hybridization of the nucleotide sequence; b. subjecting the sample and the DNA probe to stringent hybridization conditions; and c. detecting the DNA probe and the inclusion comprising a composition selected from the group consisting of SEQ ID NO: 3 or SEQ ID NO: Hybridization of DNA molecules of the nucleotide sequence of the population. 11 . A DNA detection kit comprising at least one DNA molecule comprising a nucleotide sequence having a contiguous nucleotide sequence of sufficient length of SEQ ID NO: 6 and a complement thereof for use as a specificity detector A DNA primer or probe present in the DNA of the test line 17053, wherein the detection of the DNA identifies the presence of the rice line 17053 in the sample. 12. The DNA detection kit of claim 11, wherein the DNA molecule comprises at least 15 contiguous nucleotides of SEQ ID NO: 1 or SEQ ID NO: 2 or Complement the sequence. 13. The DNA detection kit of claim 11, wherein the kit comprises a method comprising: a. contacting the sample with a DNA molecule pair as claimed in claim 7; b. performing sufficient to generate SEQ ID NO: 1 or a nucleic acid amplification reaction of an amplicon of at least 15 contiguous nucleotides of SEQ ID NO: 2 or a complement thereof; and c. detecting the amplicon. 14. A rice plant, seed or part thereof comprising line 17053, wherein 147377.doc 201040274 comprises a representative seed sample of line 17053 deposited at AT (:c accession number PTA-9843. ° 15. rice as claimed in claim 14 a plant, a seed or a part thereof, wherein the plant or a part thereof is tolerant to the treatment of a glyphosate (glyphGSate) herbicide. The rice plant or seed of claim 14, wherein the rice plant or seed has at least A hybrid comprising a parent of line 17〇53, a representative seed sample comprising line 17053 has been deposited with the ATCC registration number ρτΑ_ 9843 ° 17. A rice product produced by the seed of claim 14 wherein the commodity comprises The line is 17053. 18_ The item of claim 17 is further defined as selected from the group consisting of whole or processed rice seeds, animal feed, oil, meal, flour, sugar, popcorn (pUffed Hce), milk, milk road, paper, cream, wine 'alcohol, biomass and fuel products. 19. The rice plant part of claim 14 is defined as fine , pollen, ovule, pod, flower, root tissue, stem tissue or leaf tissue. 20. The rice plant of claim 14, further defined as a progeny plant of any generation comprising the rice plant of the line 17053, wherein the progeny The plant comprises line 17053. 21. A rice plant or a part thereof containing line 17053 capable of producing an amplicon of 疋17053. 22. The rice plant of claim 21, wherein the amplicon comprises SEq iD N〇: 1 or SEQ ID NO: 2. 147377.doc 201040274 23. A subfamily containing the line 17〇53, wherein the seed is capable of producing an amplicon of the identification line 17 0 5 3. 24. Rice seed, wherein the δ hai amplicon comprises SEQ id NO· 1 or SEQ ID NO: 2. 25. Complete or knife-free seed, animal feed, oil, meal, fine powder, debris as claimed in claim 18. , 糠, popcorn, milk, milk road, paper, milk β / 酉 ethanol &amp; material and fuel products, the further step is defined as the inclusion of a type of identification system 丨7053 when tested in the ship amplification method Increased nucleic acid, the expansion of the thumb and the human CT ^ τ expands the scorpion package 3 SEQ ID NO: 1 or SEQ ID NO. 2. 26. A method of producing a glyphosate-tolerant herbicide-tolerant rice plant comprising introducing an α-portage 1 7〇53 into the In the genome of a plant. - -27. The method of claim 26, which is defined as comprising the steps of: Μ吏 hybridizing the first rice plant comprising line 17053 with the second rice plant of the defective line 17〇53 to produce progeny Plant; and 〇b. Select at least one generation of plants comprising the line 17053 and tolerant to glyphosate. 28. The method of claim 27, further comprising self-containing the progeny plant to produce a sub-secondary plant and selecting at least one homozygous first plant of the line i 7〇53. 29. A glyphosate resistant rice plant, seed or DNA-containing portion thereof comprising a glyphosate resistant CP4 EPSPS protein and capable of producing an amplicon of the identification line 17053. A method of producing a rice product, comprising: 147377.doc 201040274 a. obtaining a rice plant or a part thereof as claimed in claim 14; and b. producing a rice product from the rice plant or a part thereof. 31. The method of claim 3, , °Hai Shang 1113 is selected from whole or processed rice seeds, animal feed, ~ one by coarse, fine powder, debris, glutinous rice, popped rice, milk, A group consisting of breast, paper, dairy, and Kongyue/west, ethanol, biomass, and fuel products. 3 2 · A method of controlling weed growth in a field comprising a rice plant of line 17 G 5 3, the method comprising treating the field with glyphosate in an amount effective to control the growth of the weed, wherein the rice plants exhibit a The grass is the right one. 33. The method of claim 32 wherein the effective amount of glyphosate is from about U pounds to about 4.5 pounds per acre. 3 = claim 29 of the glyphosate-bearing rice plant or its scorpion-containing portion, wherein the amplicon of the identification line 17053 comprises seq ι〇 or SEQ ID NO: 2. 35. A representative seed sample of an inaninal plant material comprising line 17〇53 comprising line nos] has been deposited with the ATCC accession number ρτΑ_9843, the plant material comprising selected from the group consisting of SEQ ID Ν〇: 丨, SEQ 1〇ν〇: 2 and its complementary sequence consisting of a group of dna molecules. 36. A microorganism comprising a polynucleic acid molecule having a nucleotide sequence selected from the group consisting of seq, SEQ ID 2 and the complement thereof. 37. The microorganism of claim 36, wherein the microorganism is a plant cell. 147377.doc